阅读说明：本技术 Hla-g转录物和同种型及它们的用途 (HLA-G transcripts and isoforms and their uses ) 是由 E·D·卡罗塞拉 戴安娜·特罗尼克-勒·鲁 简-菲利普·戴斯里斯 杰罗姆·韦瑞尼 弗朗索瓦· 于 2018-07-24 设计创作，主要内容包括：本发明涉及HLA-G分子及其治疗用途的领域。本发明涉及新的HLA-G同种型,即衍生自HLA-G基因的新的RNA转录物和蛋白质,包含其的药物组合物,以及这些转录物的特异性引物和这些蛋白质的特异性抗体。本发明进一步涉及这些分子的诊断或治疗用途。(The present invention relates to the field of HLA-G molecules and their therapeutic uses. The present invention relates to novel HLA-G isoforms, i.e., novel RNA transcripts and proteins derived from HLA-G genes, pharmaceutical compositions comprising the same, as well as primers specific for these transcripts and antibodies specific for these proteins. The invention further relates to the diagnostic or therapeutic use of these molecules.)

1. An isolated HLA-G protein having a sequence with at least one of the following characteristics:

-the isolated HLA-G protein comprises five amino acid residues MKTPR in its N-terminal part, i.e. SEQ ID NO:1, and/or;

-the isolated HLA-G protein lacks the α 1 domain, that is to say the sequence SEQ ID No. 3, and/or;

-the isolated HLA-G protein is free of transmembrane/cytoplasmic domains, that is to say of the sequence SEQ ID No. 6, with the proviso that the protein consists of any of the sequences SEQ ID No. 90, 91 and 92, and/or;

-said isolated HLA-G protein comprises amino acids resulting from retention of at least a portion of an intron, with the proviso that said intron is not intron 2 or intron 4.

2. The isolated HLA-G protein according to claim 1, wherein the sequence thereof comprises or consists of a sequence selected from the group consisting of SEQ ID NO 7 to SEQ ID NO 31.

3. The isolated HLA-G protein of claim 1, wherein said isolated HLA-G protein comprises 5 amino acid residues MKTPR in an N-terminal portion thereof.

4. The isolated HLA-G protein of claim 1, wherein the sequence thereof comprises or consists of a sequence selected from the group consisting of SEQ ID NOs 7, 8, 9, 10, 11, 12, 13, 14, 15, 16.

5. The isolated HLA-G protein of claim 1, wherein the sequence thereof comprises or consists of a sequence selected from the group consisting of SEQ ID NOs 9, 10, 11, 12, 13, 14, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 29, 30, 31.

6. The isolated HLA-G protein of claim 1, wherein the sequence thereof comprises or consists of a sequence selected from the group consisting of SEQ ID NOs 8, 10, 12, 14, 16, 18, 20, 21, 23, 25, 27, 28, 30.

7. An isolated polynucleotide encoding a protein as defined in any one of claims 1 to 6.

8. A recombinant vector comprising a polynucleotide as defined in claim 7.

9. A protein as defined in any one of claims 1 to 6, a polynucleotide as defined in claim 7, and/or a vector as defined in claim 8 for use as a medicament, preferably for the treatment of an autoimmune and/or inflammatory disease, or for the prevention and/or treatment of allograft rejection.

10. A protein as defined in any one of claims 1 to 6, a polynucleotide as defined in claim 7, and/or a vector as defined in claim 8 for use in the prevention and/or treatment of ischemia, preferably ischemia associated with cardiovascular disease, peripheral arterial disease or stroke.

11. A protein as defined in any one of claims 1 to 6, a polynucleotide as defined in claim 7, and/or a vector as defined in claim 8 for use in the prevention and/or treatment of vascular retinopathy.

12. An antisense oligonucleotide having a sequence complementary to at least a portion of:

-SEQ ID NO:32；

33 or-SEQ ID NO;

-SEQ ID NO:34。

13. an antibody specific for at least one of the peptides of sequences SEQ ID NO 35, 36 or 37.

14. An in vitro method for diagnosing cancer, comprising detecting in a biological sample of a subject at least an RNA transcript having a sequence selected from the list consisting of the sequences SEQ ID NOs 38 to 74 and/or at least one protein as defined in any of claims 1 to 6, preferably wherein said detection is performed at least using primers having a sequence selected from the list consisting of SEQ ID NOs 75 to 79.

15. A kit comprising at least two primers having a sequence selected from the list consisting of SEQ id nos. 78 to 84, preferably comprising at least one primer combination selected from the list consisting of:

primer Ex1F of sequence SEQ ID NO. 76 or primer PrPrPrPrPrFr of sequence SEQ ID NO. 77, and primer G526R of sequence SEQ ID NO. 80;

-primer Int4R of sequence SEQ ID No. 75 and primer Int3F of sequence SEQ ID No. 78;

primer Int3F of sequence SEQ ID No. 78 and primer Int5R of sequence SEQ ID No. 79.

Technical Field

The present invention relates to the field of HLA-G molecules and their therapeutic uses. The present invention relates to novel HLA-G isoforms, i.e., novel RNA transcripts and proteins derived from HLA-G genes, pharmaceutical compositions comprising the same, as well as primers specific for these transcripts and antibodies specific for these proteins. The invention further relates to the diagnostic or therapeutic use of these molecules.

Background

HLA-G is an HLA class Ib molecule with potent immunomodulatory activity that is expressed under physiological conditions where modulation of the immune response is required to avoid allograft recognition (i.e. maternal-fetal interface or transplant patient). HLA-G was first described to play a key role in maintaining pregnancy [1] and was found to be constitutively expressed in the fetal maternal interface in extravillous cytotrophoblast cells.

HLA-G has tolerogenic (tolerogenic) effects, modulating adaptive and innate immunity through interaction with T or B lymphocytes, NK cells or polymorphonuclear cells this effect is mediated by binding the fully soluble isoform and membrane-bound isoform directly to inhibitory receptors through the α 3 domain indeed, B and T lymphocytes of myeloid lineage, NK cells and monocytes express the immunoglobulin-like transcript ILT2(CD85j, ILIRB1) [15], monocytes, macrophages and dendritic cells express ILT-4(CD85d, LIB 2) [16] killer immunoglobulin-like receptors (KIR2DL4/p49) are specific for HLA-G and expressed by decidua NK cells unlike other inhibitory receptors which may also mediate activation [17, 18] additionally soluble HLA-G triggers apoptosis of T cells and NK cells through CD 8-like class I soluble molecules [19 ].

In normal cases, HLA-G expression is restricted to only some tissues, and in pathological cases, it is strongly increased. Indeed, HLA-G is overexpressed at high levels in several pathological conditions, including solid and hematological tumors. Over-expression of membrane-bound HLA-G and soluble HLA-G has been detected in different human solid and hematological tumors and may represent a mechanism by which tumor cells escape immune system control by inhibiting NK and T cell-mediated lysis. In particular, a high incidence of HLA-G expression has been reported in clear cell renal cell carcinoma (ccRCC), one of the most common human renal malignancies [4] [2, 3 ]. In addition, the role of HLA-G as an immune checkpoint enabling escape of tumors has been demonstrated in murine models [5, 6 ].

On the other hand, loss of HLA-G mediated control of the immune response may lead to the onset of autoimmune/inflammatory diseases caused by uncontrolled activation of immune effector cells. Several studies in recent years have shown that HLA-G plays an important role in the control of autoimmune/inflammatory diseases such as Multiple Sclerosis (MS), Crohn's Disease (CD), psoriasis, pemphigus, celiac disease, Systemic Lupus Erythematosus (SLE), asthma, juvenile idiopathic arthritis, and Rheumatoid Arthritis (RA) [23 ].

7 HLA-G isoforms have been identified, 4 of which are membrane-bound (HLA-G1, HLA-G2, HLA-G3 and HLA-G4), and 3 of which are soluble (HLA-G5, HLA-G6 and HLA-G7). All of these HLA-G include a peptide signal at their N-terminus.

HLA-G1 protein isoforms include three external domains (α 1, α 2, and α 03), a transmembrane region, and a cytoplasmic domain HLA-G2 protein isoforms do not include the α 12 domain (i.e., α 1 and α 3 domains are directly linked), followed by the transmembrane and cytoplasmic domains HLA-G3 protein isoforms lack the α 2 and α 3 domains, i.e., it includes the α 1 domain directly linked to the transmembrane and cytoplasmic domains HLA-G4 protein isoforms lack the α 3 domain, i.e., it includes the α 1, α 2, transmembrane, and cytoplasmic domains.

Soluble HLA-G isoforms all lack a transmembrane domain and a cytoplasmic domain. Interestingly, all of these soluble HLA-G proteins contain other amino acids not present in any membrane-bound HLA-G due to the retention of one intron. More specifically:

HLA-G5 protein isoforms comprise the α 1, α 2 and α 3 domains, as well as an additional C-terminal peptide sequence of 21 amino acid residues encoded by intron 4 (intron 4 remaining after transcriptional splicing and RNA maturation);

HLA-G6 protein isoform corresponds to HLA-G5 without α 2, i.e. HLA-G6 comprises the α 1 and α 3 domains, as well as an additional C-terminal peptide sequence of 21 amino acid residues encoded by intron 4 (intron 4 remaining after transcriptional splicing and RNA maturation);

HLA-G7 protein isoforms contain only the α 1 domain, and 2 additional C-terminal amino acid residues encoded by intron 2 (intron 2 remaining after transcriptional splicing and RNA maturation);

all seven reported HLA-G isoforms are derived from alternative (alternative) splicing of a primary transcript, have similar translation initiation sites, and have not proposed unique functional roles.

To date, numbering of HLA-G gene exons is based on the IMGT/HLA database (also referred to herein as IMGT/HLA nomenclature) and is described as comprising 8 exons, 7 introns, and a 3 'untranslated end, corresponding to exon 1: signal sequence, exon 2: α 1 extracellular domain, exon 3: α 2, extracellular domain, exon 4: α 3 extracellular domain, exon 5: transmembrane domain, exon 6: cytoplasmic domain I, exon 7: cytoplasmic domain II (untranslated), exon 8: cytoplasmic domain III (untranslated), and a 3' untranslated region, respectively.

However, according to the Ensembl database, the HLA-G gene may have a complementary exon at the 5' end that is not present in the IMGT/HLA database. In addition, since exon 7 corresponds to an untranslated domain, the question of whether or not it is considered to be an exon itself is of interest remains unsolved.

Thus, the presence of this complementary exon will alter the size of the 5' untranslated region (UTR) and the location of the promoter. This may alter the regulation of the gene by modifying the binding of proteins and/or mirnas.

Therapeutic approaches based on synthetic HLA-G derived proteins or antibodies have emerged in mouse models and these new therapeutic tools may prove useful for the treatment of cancer, infectious diseases, autoimmune/inflammatory diseases, and allograft rejection. Furthermore, soluble forms of HLA-G1 (also known as HLA-G5) have been shown to inhibit angiogenesis and have been proposed as therapeutic targets for the prevention of pathological neovascularization [28 ].

Therefore, in this case, a new therapeutic approach based on HLA-G molecules is required.

Disclosure of Invention

The inventors have discovered novel transcripts of the HLA-G gene, most likely due to alternative splicing.

The inventors have demonstrated the presence of HLA-G transcripts having at the 5' end a complementary sequence corresponding to the region upstream of exon 1 (according to IMGT/HLA nomenclature). Interestingly, these transcripts (referred to herein as long HLA-G transcripts) also have 106bp deletions compared to previously known HLA-G transcripts, and have ATGs that can serve as translation initiation origins [29 ].

These results confirm the hypothesis that the use of the new nomenclature based on the Ensembl database is meaningful, therefore, the Ensembl nomenclature will be used below, unless otherwise indicated, in this new nomenclature (as shown in FIG. 1) the first exon is located within the 5 'terminal complement sequence, while the previous exon 7 has been deleted, thus, the exon numbering is thus modified in that exon 1: corresponds to the newly found sequence, exon 2: signal sequence, exon 3: α 1 extracellular domain, exon 4: α 2 extracellular domain, exon 5: α 3 extracellular domain, exon 6: transmembrane domain, exon 7: cytoplasmic domain I, exon 8: cytoplasmic domain III (untranslated) and 3' untranslated regions (compare IMGT/HLA nomenclature, exon 1 is renamed exon 2, exon 2 is renamed exon 3, etc.).

Furthermore, the inventors have discovered novel HLA-G transcripts which retain intron 1, 4, 6 or 7, and transcripts which retain both introns (particularly intron 3 and 4 or intron 3 and 5), which have never been reported before.

The inventors further showed that alternative splicing makes it possible for a new translation initiation codon to be present, which is different from the one located in exon 2, which was considered to be the only possible translation initiation codon for HLA-G protein to date. In the novel transcript, possible translation initiation codons were found in exon 1 and exon 4.

In other words, the novel transcripts encode novel HLA-G proteins each having a structure different from HLA-G isoforms that have been disclosed so far.

Among these novel transcripts, the inventors have identified some major features related to HLA-G proteins that have not been previously disclosed.

The first feature is that in some of the novel HLA-G proteins identified, there are five amino acid residues MKTPR at their N-terminus, i.e. upstream of exon 1 (according to IMGT/HLA nomenclature). This feature results from the initiation of translation in exon 1. For convenience and clarity only, and regardless of the actual length of their amino acid sequences, HLA-G proteins having this characteristic are referred to herein as "long HLA-G" to highlight the presence of the additional amino acid residues.

The second feature is the absence of the α 1 domain in some of the novel HLA-G proteins identified.

The third feature is the absence of transmembrane domains in some of the novel HLA-G proteins identified, which have peptide sequences different from the known soluble HLA-G proteins HLA-G5, HLA-G6 and HLA-G7. For convenience and clarity only, HLA-G proteins having these characteristics are referred to herein as "soluble HLA-G".

A fourth feature is that, in some of the novel HLA-G proteins identified, at least a portion of the intron is retained differently from intron 2 or 4 (such retention is observed in HLA-G5, HLA-G6 and HLA-G7). All newly identified HLA-G proteins are expected to be tolerogenic, consistent with the reported function of known HLA-G proteins. Thus, these proteins are useful in the treatment of autoimmune/inflammatory diseases such as Multiple Sclerosis (MS), Crohn's Disease (CD), psoriasis, pemphigus, celiac disease, Systemic Lupus Erythematosus (SLE), asthma, juvenile idiopathic arthritis, and Rheumatoid Arthritis (RA), as well as in the prevention of allograft rejection.

In addition, based on the knowledge in the art, the newly discovered HLA-G transcripts and corresponding proteins are expected to play a role in the ability of cancer cells to evade immune checkpoints and thus constitute important therapeutic targets. In this context, the inventors have designed molecules targeting novel RNA transcripts or their encoded proteins, as well as compositions comprising such molecules, which are useful for the treatment of cancer, in particular for the treatment of clear cell renal cell carcinoma (ccRCC).

As detailed in the experimental section, the inventors have demonstrated that newly identified HLA-G proteins have a surprising angiogenic effect in vivo. This effect is in sharp contrast to the effect reported in the literature for HLA-G5/soluble HLA-G1. In one aspect, the newly identified HLA-G proteins may therefore be useful in the treatment of conditions requiring angiogenesis (such as ischemia, which is a symptom found, for example, in cardiovascular disease, peripheral arterial disease, and stroke) due to therapeutic angiogenesis.

Further, the inventors designed primers and antibodies that can be used to detect these transcripts and their encoded proteins, and thus can be used to diagnose cancer. Accordingly, a first aspect of the invention is an isolated HLA-G protein, the sequence of which has at least one of the following characteristics:

-it comprises five amino acid residues MKTPR in its N-terminal part, i.e. SEQ ID NO:1, and/or;

it has NO α 1 domain, that is to say NO sequence SEQ ID NO 3, and/or;

it has NO transmembrane/cytoplasmic domain, that is to say NO sequence SEQ ID NO 6, and;

it includes amino acids resulting from the retention of at least part of one intron, with the proviso that (proviso) said intron is not intron 2 or intron 4.

The term "HLA-G protein" encompasses any protein or polypeptide resulting from the expression of an HLA-G gene, preferably the human HLA-G gene of gene ID 3135 (as mentioned in the GeneBank database based on the genomic reference GRCh38. p10.) preferably, an "HLA-G protein" is a protein or polypeptide comprising a sequence corresponding at least to the translation of any of exons 1 to 8 of a human HLA-G gene according to Ensembl nomenclature.

In the context of the present invention, a "peptide signal" has the sequence SEQ ID NO 2.

In the context of the present invention, the "α 1 domain" has the sequence SEQ ID NO 3.

In the context of the present invention, the "α 2 domain" has the sequence SEQ ID NO 4.

In the context of the present invention, the "α 3 domain" has the sequence SEQ ID NO 5.

In the context of the present invention, the "transmembrane/cytoplasmic domain" has the sequence SEQ ID NO 6.

The term "amino acids resulting from intron retention" should be interpreted as commonly understood in the art.in the context of the present invention, a person skilled in the art can readily identify such amino acids by comparing the sequence of the protein with a reference sequence without any amino acids resulting from intron retention, such as a reference sequence consisting of translation of all exons of the HLA-G gene.A reference sequence suitable for such comparison is the sequence SEQ ID NO:7, which corresponds to the peptide sequence of an HLA-G protein having all exons of the HLA-G gene and without introns, that is to say having five amino acid residues MKTPR, peptide signals, α 1, α 2 and α 3 domains, transmembrane/cytoplasmic domains in its N-terminal part.

Preferably, in the context of the present invention, a "sequence of an HLA-G protein without transmembrane/cytoplasmic domain, that is to say a sequence without the sequence SEQ ID NO 6" is a protein consisting of:

-at least a portion of SEQ ID NO 1; and/or;

-at least a portion of SEQ ID No. 2; and/or;

-at least a portion of SEQ ID No. 3; and/or;

-at least a portion of SEQ ID No. 4; and/or;

-at least a portion of SEQ ID No. 5; and

with the proviso that the protein does not have the sequence of HLA-G5, HLA-G6 or HLA-G7, that is to say consists of any of the sequences of SEQ ID NO:90, 91 and 92, respectively.

The inventors have specifically identified several novel HLA-G proteins according to the present invention. The isolated HLA-G protein according to the present invention is preferably a protein whose sequence comprises or consists of a sequence selected from the group consisting of SEQ ID NO 7 to SEQ ID NO 31.

Preferably, the HLA-G protein according to the present invention (the sequence comprising five amino acid residues MKTPR in its N-terminal part) is a protein whose sequence comprises or consists of a sequence selected from the group consisting of SEQ ID NOs 7, 8, 9, 10, 11, 12, 13, 14, 15, 16.

Preferably, the HLA-G protein according to the invention without the sequence of the α 1 domain is a protein whose sequence comprises or consists of a sequence selected from the group consisting of SEQ ID NOs 9, 10, 11, 12, 13, 14, 17, 18, 19, 20, 22, 23, 24, 25, 26, 27, 29, 30, 31.

Preferably, the HLA-G protein according to the invention without the sequence of the α 2 domain is a protein whose sequence comprises or consists of a sequence selected from the group consisting of SEQ id NOs 8, 10, 12, 13, 14, 16, 18, 20, 21, 23, 25, 27, 28, 30.

The HLA-G protein according to the present invention may be a purified protein or a synthetic protein, which can be obtained by conventional techniques known to those skilled in the art.

Preferably, the invention also includes variants of the above proteins, that is to say proteins whose sequence has at least 80, 85, 90 or 95% identity with at least one of the above proteins.

In the sense of the present invention, "percent identity" or "% identity" between two nucleic acid or amino acid sequences refers to the percentage of identical nucleotides or amino acid residues between the two sequences to be compared, obtained after optimal alignment, this percentage being purely statistical, and the differences between the two sequences being randomly distributed along their length. Traditionally, two nucleic acid or amino acid sequences are compared after optimal alignment of the sequences, either in segments or using an "alignment window". In addition to manual comparisons, comparisons for optimal alignment of sequences can be made by the local homology algorithms of Smith and Waterman (1981), by the similarity search method of Pearson and Lipman (1988), or by methods of computer software using these algorithms (GAP, BESTFIT, FASTA and TFASTA in the Wisconsin genetics software package, 575Science Dr., Madison, Wisconsin, or by the comparison software BLAST NR or BLAST P).

The percent identity between two nucleic acid or amino acid sequences is determined by comparing the two optimally aligned sequences, where the nucleic acid or amino acid sequences to be compared may have additions or deletions compared to the reference sequence, to achieve optimal alignment between the two sequences. Percent identity is calculated by determining the number of positions at which the amino acids, nucleotides or residues between two sequences, preferably between two complete sequences, are identical, dividing the number of identities by the total number of positions in the alignment window, and multiplying the result by 100 to obtain the percent identity between the two sequences.

For example, default parameters (particularly for the parameters "open gap penalty": 5, and "extended gap penalty": 2; the selected matrix is, for example, the program set forth "BLOSUM 62" matrix) may be used with the BLAST program "BLAST 2 sequences" [27] (available from the http:// www.ncbi.nlm.nih.gov/gorf/bl2.html website); the percent identity between the two sequences to be compared is directly calculated by the program.

Preferably, the present invention further comprises functional variants of HLA-G proteins according to the present invention, wherein the amino acid residues are substituted with other amino acid residues having similar chemical properties (e.g. charge or hydrophobicity) and thus do not alter the functional properties of the molecule.

Preferably, the present invention also includes modified proteins derived from the above proteins by introducing any chemical modification into one or more amino acid residues, peptide bonds, N-and/or C-termini of the proteins, with the aim of increasing the stability, bioavailability or bioactivity of the protein, as long as the modified protein retains function.

As is well known in the art, an alternative to using an isolated protein in vivo may be to use a polynucleotide encoding the protein in an expressible form or a recombinant vector comprising the polynucleotide. Such vectors may be useful for the production of proteins or for in vivo therapeutic use.

Another aspect of the invention relates to isolated polynucleotides encoding the proteins of the invention. The synthetic or recombinant polynucleotide may be single-and/or double-stranded DNA, RNA, or a combination thereof. Preferably, the polynucleotide comprises a coding sequence optimized for the host in which the protein is expressed.

Another aspect of the invention relates to a recombinant vector comprising said polynucleotide. Preferably, the recombinant vector is an expression vector capable of expressing the polynucleotide when transfected or transformed into a host cell. The polynucleotide is inserted into an expression vector in the proper orientation and correct reading frame for expression. Preferably, the polynucleotide is operably linked to at least one transcriptional regulatory sequence, and optionally to at least one translational regulatory sequence. Recombinant vectors include commonly used vectors for genetic engineering and gene therapy, including, for example, plasmids and viral vectors.

Another aspect of the invention provides a host cell transformed with the polynucleotide or recombinant vector.

The polynucleotides, vectors and/or cells of the invention may be prepared using well known recombinant DNA techniques.

The proteins of the invention are expected to have tolerability similar to HLA-G proteins known in the art, particularly since all newly discovered HLA-G proteins disclosed herein have the α 3 domain.

Another aspect of the invention relates to a pharmaceutical composition comprising at least one protein, polynucleotide and/or vector of the invention, and preferably a pharmaceutically acceptable carrier.

Suitable vehicles or carriers include any pharmaceutically acceptable vehicle, such as buffers, stabilizers, diluents, salts, preservatives, emulsifiers, sweeteners, and the like. The vehicle typically comprises an isotonic aqueous or non-aqueous solution, which may be prepared according to known techniques. Suitable solutions include buffered solutes such as phosphate buffer solutions, chloride solutions, ringer's solutions, and the like.

Another aspect of the invention relates to a protein, polynucleotide and/or vector of the invention for use as a medicament.

Preferably, the above-described proteins, polynucleotides and/or vectors of the invention are used for the treatment of autoimmune and/or inflammatory diseases, or for the prevention and/or treatment of allograft rejection, for the prevention and/or treatment of ischemia (preferably ischemia associated with cardiovascular disease, peripheral arterial disease or stroke), or for the prevention and/or treatment of vascular retinopathy.

In the context of the present invention, the term vascular retinopathy includes von hippel angiomatosis, cavernous retinal hemangioma, exudative retinopathy, idiopathic macular telangiectasia (rices disease), obstruction of the central retinal artery, obstruction of the arterial branches, obstruction of the ciliary retinal arteries, obstruction of the ocular arteries, retinal vein obstruction.

In the context of medical use, a person skilled in the art can preferably select a protein having the α -3 domain in the HLA-G protein of the invention, such as any of the proteins of the sequences SEQ ID NO:7, 8, 9, 10, 13, 14, 17, 18, 19, 20, 21, 22, 23, 26, 27, 31.

Advantageously, the HLA-G protein for use as a medicament, preferably for use in the above treatment, is a protein having a sequence comprising or consisting of a sequence selected from the group consisting of SEQ ID NO 7, 8, 9, 10, 13, 14, 17, 18, 19, 20, 21, 22, 23, 26, 27, and 31.

In the context of the present invention, autoimmune/inflammatory diseases preferably mean Multiple Sclerosis (MS), Crohn's Disease (CD), psoriasis, pemphigus, celiac disease, Systemic Lupus Erythematosus (SLE), asthma, juvenile idiopathic arthritis, and Rheumatoid Arthritis (RA), but preferably psoriasis.

The present invention also provides a method for the treatment of autoimmune/inflammatory diseases, or the prevention and treatment of allograft rejection, for the prevention and/or treatment of ischemia (preferably ischemia associated with cardiovascular disease, peripheral arterial disease or stroke), or for the prevention and/or treatment of vascular retinopathy, comprising: administering to the individual a therapeutically effective amount of at least one protein, polynucleotide and/or vector of the invention, or a composition as described above.

By "therapeutically effective amount" is meant herein an amount sufficient to at least reduce or prevent the progression of the disease over time. Generally, the amount can be adjusted by the skilled artisan according to the pathological condition, the subject, the duration of the treatment, the presence of other active ingredients, and the like.

As already said, the newly discovered HLA-G transcripts and the corresponding proteins are important therapeutic targets, in particular for the treatment of cancer. Typically, for therapeutic purposes, RNA transcripts can be targeted by antisense oligonucleotides, while proteins can be targeted by specific antibodies.

For example, an RNA transcript that does not contain the sequence encoding the α 1 domain but contains the sequences encoding the signal peptide and the α 2 and α 3 domains, with a linkage between the sequence encoding the signal peptide and the sequence encoding the α 2 domain, corresponding to the sequence SEQ ID NO:32 that is not found in other HLA-G proteins similarly, an RNA transcript consisting of the sequence encoding the α 3 domain and the signal peptide but not the α 1 and α 2 domains includes the sequence SEQ ID NO:33, which corresponds to the linkage between the sequence encoding the α 3 domain and the signal peptide, an RNA transcript encoding soluble HLA-G includes the sequence SEQ ID NO: 34.

Another object of the invention is an antisense oligonucleotide having a sequence complementary to at least a portion of:

-SEQ ID NO:32；

33 or-SEQ ID NO;

-SEQ ID NO:34。

the invention further relates to a recombinant vector comprising a polynucleotide encoding at least one antisense oligonucleotide of the invention.

The novel HLA-G proteins of the present invention have specific peptide sequences SEQ ID NO 35, 36 and 37 encoded by the linker sequences SEQ ID NO 32, 33 and 34, respectively, which can be used as antigens to generate antibodies specific to such proteins, i.e., anti-HLA-G antibodies specific to certain HLA-G proteins.

Another object of the invention is an antibody specific for at least one peptide of sequence SEQ ID NO 35, 36 or 37.

For the purposes of the present invention, the term "antibody" refers to an immunoglobulin that specifically binds to another molecule and is therefore defined as complementary to a particular spatial and polar organization of another molecule. The antibody may be a monoclonal antibody or a polyclonal antibody, and may be prepared by techniques well known in the art, such as immunizing a host and collecting serum (polyclonal), or preparing a continuous hybrid cell line and collecting secreted protein (monoclonal), or by cloning and expressing a nucleotide sequence encoding at least the amino acid sequence required for specific binding of a native antibody, or a mutagenized version thereof. Antibodies may include whole immunoglobulins or fragments thereof, which include various classes and isotypes, such as IgA, IgD, IgE, IgG1, IgG2a, IgG2b, and IgG3, IgM. Functional antibody fragments can include portions of an antibody (e.g., Fab, Fv, and F (ab ')2 or Fab') that are capable of retaining binding with similar affinity as a full-length antibody. In addition, aggregates, polymers and conjugates of immunoglobulins or fragments thereof may be used where appropriate, so long as the binding affinity for the particular molecule is substantially maintained.

The term "antibody specificity of at least one peptide" should be construed as generally understood in the art, that is, to indicate that the antibody exhibits a significant affinity for the at least one peptide, preferably about 10^-5M (KD) or higher. Affinity can be determined by a variety of methods well known to those skilled in the art, including but not limited to Biacore analysis, Blitz analysis, and Scatchard plots.

Another object of the invention is a pharmaceutical composition comprising an antibody, an antisense oligonucleotide and/or a vector encoding same according to the invention, and preferably a pharmaceutically acceptable carrier.

Another aspect of the invention relates to the antibody, antisense oligonucleotide, and/or vector encoding the same of the invention, for use as a medicament.

Preferably, the above-described antibody, antisense oligonucleotide and/or vector encoding the same of the present invention are used for treating cancer.

Non-limiting examples of cancer include esophageal cancer, gastric cancer, colon cancer, pancreatic cancer, melanoma, thyroid cancer, lung cancer, breast cancer, kidney cancer, bladder cancer, uterine cancer, ovarian cancer, and prostate cancer; hepatocellular carcinoma, osteosarcoma, cylindroma, neuroblastoma, glioblastoma, astrocytoma, colitis-related carcinoma, multiple myeloma, and various types of leukemia and lymphomas (such as diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma (PMBL), hodgkin's lymphoma, and MALT lymphoma). In a preferred embodiment, the cancer is clear cell renal cell carcinoma (ccRCC).

The present invention also provides a method for treating cancer, the method comprising: administering to the individual a therapeutically effective amount of an antibody, antisense oligonucleotide and/or vector encoding the same, or a composition as described above, of the invention.

As already noted, the inventors have identified that cancer cell samples include specific HLA-G transcripts, i.e., RNA molecules corresponding to human HLA-G gene expression. These RNA transcripts correspond to the sequences SEQ ID NO:38 to 74. Thus, based on the presence of these RNA transcripts in biological samples, the inventors have developed in vitro methods for diagnosing cancer.

The present invention further relates to an in vitro method for diagnosing cancer comprising detecting at least an RNA transcript having a sequence selected from the list consisting of the sequences SEQ ID NO:38 to 74 and/or at least one protein of the invention as disclosed above in a biological sample of a subject.

The term "biological sample" refers to a sample obtained from a subject, including samples of biological tissue or liquid origin. These samples can be, but are not limited to, bodily fluids (e.g., blood, plasma, serum, or urine), organs, tissues, fractions, and cells isolated from mammals including humans. The biological sample may also include a portion of the biological sample that includes a tissue (e.g., a cross-sectional portion of an organ or tissue). The biological sample may also include an extract from the biological sample, such as an antigen from a biological fluid (e.g., blood or urine). Preferably, the subject is a mammal, but preferably a human.

In the context of the present invention, detection of at least one RNA transcript having a sequence selected from the list consisting of sequences SEQ ID NO:38 to 74, and/or at least one protein of the present invention, indicates that the subject has cancer.

Non-limiting examples of cancer include esophageal cancer, gastric cancer, colon cancer, pancreatic cancer, melanoma, thyroid cancer, lung cancer, breast cancer, kidney cancer, bladder cancer, uterine cancer, ovarian cancer, and prostate cancer; hepatocellular carcinoma, osteosarcoma, cylindroma, neuroblastoma, glioblastoma, astrocytoma, colitis-related carcinoma, multiple myeloma, and various types of leukemia and lymphomas (such as diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma (PMBL), hodgkin's lymphoma, and MALT lymphoma). In a preferred embodiment, the cancer is clear cell renal cell carcinoma (ccRCC).

As described above, the detection of at least one protein of the invention can be carried out by subjecting at least one antibody specific for said protein to a suitable immunological method (e.g., ELISA, RIA, immunofluorescence, immunohistochemistry).

Detection of RNA transcripts can be carried out by hybridization of nucleotide probes specific for the RNA (e.g., attached to a biochip), or by amplification of nucleotide primers specific for the RNA transcript (e.g., by RT-PCR).

The inventors have developed nucleotide primers particularly suitable for detecting at least one RNA transcript of the invention:

primer int4R of sequence SEQ ID NO 75, designed to hybridize with the start (beginning) of intron 4 and thus can be used to detect HLA-G transcripts retaining this intron;

76, designed to hybridize with a region complementary to exon 1 and thus can be used to detect transcripts encoding long HLA-G proteins;

77 designed to hybridize with a region further upstream of the region annotated by the 5' transcription end of the gene (reported as the promoter region) and thus can be used for the detection of transcripts encoding long HLA-G proteins;

78 designed to hybridize with a region within intron 3 and which can be used to detect HLA-G transcripts retaining this intron;

79, designed to hybridize to a region within intron 5 and can be used to detect HLA-G transcripts retaining this intron;

these primers are particularly useful for carrying out the diagnostic methods of the invention. Thus, in one embodiment, in the method of the invention, the detection of at least the RNA transcript of the invention is carried out using at least a primer having a sequence selected from the list consisting of SEQ ID NO:75 to 79.

These specific primers can be used in combination with each other or in combination with other known primers specific for HLA-G transcripts, such as primer G526R of sequence SEQ ID NO:80, as already disclosed in the art. Further, any combination of primer Int3F and primer Int5R or primer Int4R can be used to detect transcripts that retain two introns ( intron 3 and 4, or intron 3 and 5).

In a preferred embodiment, in the diagnostic method of the invention, the detection of at least the RNA transcript of the invention is carried out using at least one of the following combinations:

primer Ex1F of sequence SEQ ID NO. 76 or primer PrPrPrPrPrFr of sequence SEQ ID NO. 77, and primer G526R of sequence SEQ ID NO. 80;

-primer Int4R of sequence SEQ ID No. 75 and primer Int3F of sequence SEQ ID No. 78;

-primer Int3F of sequence SEQ ID No. 78 and primer Int5R of sequence SEQ ID No. 79;

thus, another object of the invention is a primer having a sequence selected from the list consisting of SEQ ID NO 75 to 79.

The invention further relates to a kit comprising at least two primers having a sequence selected from the list consisting of SEQ id nos 75 to 80.

Preferably, the kit comprises at least a primer combination selected from the list consisting of:

primer Ex1F of sequence SEQ ID NO. 76 or primer PrPrPrPrPrFr of sequence SEQ ID NO. 77, and primer G526R of sequence SEQ ID NO. 80;

-primer Int4R of sequence SEQ ID No. 75 and primer Int3F of sequence SEQ ID No. 78;

-primer Int3F of sequence SEQ ID No. 78 and primer Int5R of sequence SEQ ID No. 79;

the invention also includes other provisions which will emerge from the following implementation examples and which should not be construed as limiting the scope of the invention.

Drawings

FIG. 1 is a schematic view of the structure of HLA-G gene. A. IMGT/HLA nomenclature (top) and Ensembl database (bottom). The numbers represent exons, and the domains of HLA-G protein are shown below. TM: across the membrane; CT: the cytoplasmic tail. B. Localization of primers for different RT-PCR strategies. The size (bp) and translation initiation codon of a particular amplicon are shown.

Figure 2-differential morphology and HLA-G staining pattern of 8 ccrccs included in this study. Trophoblast tissue was used as a positive control for immunohistochemical studies (H & E and immunoperoxidase staining).

Figure 3-expression of HLA-G1 in ccRCC patients. RT-PCR was performed on RNA using HLA-G1-specific primers G257F and G526R (upper panel) and ACTB primer as controls (lower panel). Lane 1: except for the tumors of patients 6 and 8, adjacent non-tumor regions. Lanes 2, 3 and 4: different tumor areas. For patients 6 and 8, all regions shown correspond to tumor regions since a partial nephrectomy was performed, while adjacent tumor regions were not available. M: 100bp sized marker.

FIG. 4-intron retention events found in HLA-G transcripts. Only read lengths across intron-exon junctions were considered. The read length corresponding to only the intron sequence is discarded.

Figure 5-molecular validation of major intron retention events. A: schematic representation of RT-PCR strategies developed for amplification of retained introns. B: results of RT-PCR analysis using actin primers as a control for the absence of genomic DNA (left), and Int1 and G257R primers to detect the presence of intron 1 (right). A523 bp band indicated no intron 2, and the presence of intron 2 resulted in a 649bp band. C: HLA-G transcripts of intron 4 were retained only (left panel) or several introns were retained simultaneously (middle and right panels).

FIG. 6-identification of 5' extended transcript HLA-G1. A. DNA sequence details show a shortened distance between the two ATGs. The sequence was performed up using the G526R primer. B. Schematic representation of a 106bp deletion; the two ATGs are underlined.

FIG. 7: a: photographs of NSG mice xenografted with RCC7 cells infected with a vector encoding long HLA-G1 virus taken on day 38 post injection. B: photographs of NSG mice xenografted with RCC7 cells infected with a vector encoding long HLA-G1L virus taken on day 38 post injection.

FIG. 8: a: photo of nude mice xenografted with RCC7 cells expressing GFP, HLA-G1 or HLA-G1L at day 25 after intradermal injection. B: photographs of nude mice xenografted with RCC7 cells grown on basement membrane matrix and expressing GFP, HLA-G1 or HLA-G1L on day 25 after intradermal injection.

FIG. 9: photographs of NSG immunodeficient mice were taken 8 days after left ear injection of control RCC7 cells (expressing GFP) and right ear RCC7 cells expressing HLA-G1L (a and B).

FIG. 10: photographs of NSG immunodeficient mice 8 days after left ear injection of control RCC7 cells and right ear HLA-G1 expressing RCC7 cells (a and B).

Detailed Description

A.Detection and analysis of novel HLA-G isoforms

1. Materials and methods

1.1 tumors and patients

All patients in this study received ccRCC radical nephrectomy as the first therapeutic intervention in urology in the holy lous hospital (paris, france) from 11 months 2014 to 4 months 2015. Median tumor size was 50 mm (ranging from 35 to 175). These tumors were classified as pT1a (patient 6), pT1b ( patients 1, 3 and 8), and pT3a ( patients 2, 4, 5 and 7) according to the 2010 primary tumor TNM classification. Two patients (patients 2 and 4) developed visceral metastases. Based on the world health organization's classification of kidney tumors, experienced uropathologists attribute all of these kidney tumors to ccRCC [8 ]. All patients enrolled in the study were given free and informed written consent. The study was approved by the institutional review board of the san louse hospital, paris.

1.2 tumor sample treatment

For each tumor, we isolated 3 to 10 samples of 10 × 5 × 5mm, depending on tumor size, representing spatial extent and macroscopic intratumoral heterogeneity. Half of each sample was snap frozen in liquid nitrogen within 1 hour after the renal artery was occluded and the other half was used for histological analysis and recorded by photography. The area not containing tumor cells in the histopathological examination was also isolated as a control.

1.3 immunohistochemistry

Immunohistochemical studies were performed on formalin-fixed and paraffin-embedded tumor tissue sections 4 μm thick for each tumor using murine antibodies 4H84, IgG1 (dilution 1/200, santa cruz biotechnology, santa cruz, ca) that recognize epitopes located in the α 1 domain common to all HLA-G isotypes, and two antibodies 5A6G7 and 2a12 (dilution 1/100, Exbio antibodies, Exbio co, CR) that recognize epitopes encoded by intron 5(Ensembl database) retained in soluble HLA-G5 and-G6 isotypes, using the OptiView DAB c detection kit (Roche), under short or standard antigen recovery conditions, staining was performed on sections of Roche (BenchMark ULTRA system, atlas, tusson) automated microtome, antibody was incubated at 37 ℃ for 32 minutes, counterstaining procedure for 4 minutes, and counterstaining for appropriate controls.

Immunohistochemical analysis was performed by a urologist using a BX51 microscope (Olympus France s.a.s, Rungis). Each immunostaining was scored by membrane-and/or cytoplasmic-based staining with negative, weak, medium or strong staining intensity, and negative staining distribution (0% of tumor area), minimal (0-10% of tumor), focal (< 50% tumor area) or diffuse (> 50% tumor area), respectively. Each run, trophoblast tissue was used as a positive control, and isotype-specific immunoglobulin was used as a negative control.

1.4 preparation of trophoblast cell samples

Trophoblast cell tissue is obtained from miscarriage (less than three months of pregnancy). After mechanical dissociation, the samples were stored at-80 ℃ in Trizol^TMIn the reagent (LifeTech, ref.15596-026), until RNA was extracted using the protocol described below.

1.5, RNA extraction

From in Trizol^TMTotal RNA was isolated from manually crushed tissue sections in reagent (Life technology, ref.15596026). After chloroform separation, RNA was purified using the miRNeasy mini Kit (Qiagen, ref.217004) according to the manufacturer's instructions and additionally subjected to DNase treatment (Qiagen, ref.79254). RNA purity and concentration were assessed using a Nanodrop spectrophotometer and Agilent2100 bioanalyzer system. RNA Integrity Number (RIN) value is mostly>8。

1.6、RT-PCR

RNA was reverse transcribed into cDNA using the GoScript reverse transcriptase kit (Promega, ref. a5001) and a thermal cycler Eppendorf (MasterCycler, Pro S). PCR reactions were performed using ampliTaq polymerase from LifeTech (ref. n80800166) in a final volume of 10 μ Ι _ with 2 μ Ι _ of cDNA template. For amplification, 40 cycles (30 seconds at 94 ℃, 30 seconds at 55 or 60 ℃, and 30 seconds at 72 ℃) were performed. HLA-G and Actin (ATCB) primers are described in Table 1. ATCB amplification was used as a control in all experiments. The PCR amplification products were mixed with 6X loading dye (Promega, ref. G1881) and analyzed on 100mL of 2% agarose gel stained with 2. mu.L of ethidium bromide at a concentration of 1 mg/mL. The molecular weight markers used were 1Kb plus DNA ladder from Invitrogen (ref.10787018). Imaging was performed using a ChemiDoc XRS system (Biorad) and interpreted using ImageLab software (Biorad).

TABLE 1 PCR primers for RT-PCR experiments

1.7 RNA sequencing

An index complementary DNA library was prepared from 1. mu.g of total RNA according to the Illumina TRUSEQ protocol. The average size of the purified PCR products was 275bp using AMPure XP beads (Beckman Coulter, Inc.). Paired-end 150bp read length sequencing of the transcriptome was performed in an equimolar pool of four cDNA libraries on NextSeq 500 (ILLUMINA).

1.8 high throughput analysis of HLA-G isoforms

The Ensembl nomenclature will be used throughout. Short reads from NGS sequencing were mapped to the human reference genome NCBI Hg19 using a BWA aligner (BWA MEM option) [20 ]. Low quality mapped reads were filtered out from the alignment file and samtools were used to extract reads mapped to HLA-G loci (Li et al, 2009). The intron retention test was performed by selecting reads that overlapped one of the intron and the surrounding exon, and the retention of the intron was only assessed when we detected reads that overlapped the 5 'and 3' flanking exons. Exon skipping detection is performed by analyzing read lengths that present a split map, searching for discontinuities in the order of mapped exons, for example: reads mapped to the end of exon 4 and the beginning of exon 6 but not to exon 5 indicate skipping of exon 5. Each subset of read lengths was visually verified by IGV [22 ]. For retention of intron n, the percentage of read length pni supporting this event was calculated as the ratio between the read length supporting the event (read length at exon n/intron junction, read length for internal intron to intron n, and read length at intron n/exon n +1 junction) and the total number of read lengths across the region where the event occurred (region from the junction between exon n and intron n to the junction between intron n and exon n + 1): let R (i) be the number of read lengths strictly in region i (read lengths are only in region i and do not overlap with other regions), and R (i, j) be the number of read lengths that overlap with regions i and j. Let S (I) be the number of reads that support exon I skipping (reads where exon n overlaps with exon m, where m > n + 1). Thus, the number of reads supporting intron retention is IRn ═ R (exon n, intron n) + R (intron n, exon n + 1). The intron retention region has a read length of Tn ═ IRn + R (exon, exon +1) + s (n); therefore, pni is given by pni ═ IRn/Tn.

For exon n skipping, the percentage of read length pne supporting this event is given by pne ═ s (n)/Tn. Analysis of potential deviations was evaluated by using a TopHat2 calibrator [24 ].

2. Results

2.1, apparent subcellular heterogeneity of HLA-G isoform distribution in ccRCC

To consider HLA-G as a potential target for cancer therapy, the expression of HLA-G in tumor cells derived from patients with ccRCC was evaluated, to this end, each tumor was isolated in 3 to 10 sections based on tumor size microscope analysis of hematoxylin and eosin (H & E) stained slides confirmed the morphological heterogeneity classically associated with ccRCC (FIG. 2, left panel) [8] we further dissected this heterogeneity by immunostaining with specific antibodies to HLA-G, 4H84, which recognizes an epitope located in the α 1 domain common to all seven reported HLA-G isoforms, and antibody 5A6G7, which recognizes only soluble HLA-G5 and HLA-G6 isoforms, the retained intron 5 (previously referred to as intron 4 according to IMGT/HLA nomenclature) encoding amino acids, trophoblast cells expressing HLA-G at high levels were used as a positive control.

Even if all tumors express HLA-G in at least one region, the expression is different between and within the tumors. The tumors of patients 1 and 2 showed strong immunostaining with the 4H84 antibody in all areas. Staining was membranous and cytoplasmic (fig. 2). Notably, an additional very strong staining of the hyaline globules located in the tumor cell cytoplasm was also detected. These transparent beads are clearly visible on H & E slides and constitute a very rare aspect of the tumor [9 ]. On the other hand, using the 5A6G7 antibody, weak or moderate granular cytoplasmic immunostaining was found in the cytoplasm, but not in the clearing sphere. HLA-G expression in tumors of other patients varies widely: the tumors of patients 6 and 7 showed diffuse but moderate 4H84 antibody membrane immunostaining. Immunostaining with 5A6G7 showed no (patient 6) or weak and focal (patient 7), indicating the absence of soluble proteins HLA-G5 and HLA-G6. In the other two tumors (patients 4 and 5), the expression of HLA-G as assessed by the 4H84 antibody was observed under a small microscope of only one tumor area. Notably, the only HLA-G positive area in patient 4 tumor corresponded exactly to the intracytoplasmic clear pellet. No staining was observed in any other area of the tumor.

The immunostaining patterns of tumor cells of patients 3 and 8 were unexpected.no immunostaining was detected with 4H84 antibody labeling all reported HLA-G isoforms the lack of tumor sections labeled with this antibody can usually be interpreted as no HLA-G expression however, diffuse strong granular intracytoplasmic 5A6G7 immunostaining, as well as diffuse, thin and granular intracytoplasmic immunostaining are observed in tumor cells of patients 3 and 8 respectively, this is unpredictable considering our current understanding of the structure of the seven HLA-G isoforms reported, as they both contain the α 1 domain recognized by the 4H84 antibody, to try to better understand these differences we have performed similar analyses using the antibody named 2a12, which also recognizes the epitope encoded by the retained intron 5 (ensel database) present in HLA-G5 and-G6 isoforms, the results show different and unexpected immunostaining patterns, especially in the clear-labeled spherules of patients 1 and 2.

In summary, the results of immunohistochemical studies clearly indicate intra-and inter-heterogeneity of HLA-G expression in ccRCC tumors. However, certain immunostaining patterns are unexpected insofar as our general understanding of the structure of HLA-G isoforms is concerned.

2.2 investigation of HLA-G1 transcript expressed in ccRCC

To better understand the HLA-G isoforms expressed in ccRCC and to elucidate the results of immunohistochemical analysis, HLA-G isoform diversity was further assessed by RT-PCR. Tumor sections from the eight patients studied above were amplified using the well-known primers G257F and G526R shown schematically in FIG. 1B [10 ]. These primers amplify a region containing the epitope recognized by the 4H84 antibody. Actin mRNA amplification was performed for each sample as a control. A predetermined 290bp band was found in all tumor sections from patients 1, 2 and 6, which band was specific for amplification of HLA-G1 transcript, and was detected in only one or two regions of the tumors from other patients (FIG. 3). No amplification products were detected in non-tumor adjacent tissues. Since the sequences of the different isoforms are highly similar and these RT-PCR conditions do not allow identification of other isoforms lacking exon 4 (the target of primer G526) (e.g., HLA-G2, -G3, -G6 or-G7), we performed a large scale study of RNAseq with the aim of providing a comprehensive description of the isoforms expressed in ccRCC.

2.3, RNA-Seq shows unannotated HLA-G transcripts

The RNAseq technique offers the strongest approach to analysis of expressed isoforms, offering the opportunity to detect alternative splicing events and unannotated transcripts, which is crucial for understanding species development and disease mechanisms [25 ].

At first sight, we have sequenced four representative samples at very high depth of coverage (depth >300 ×). Read lengths were aligned and quantified according to the Ensembl 70(GRCh37.p8) reference annotation described in materials and methods. Alternatively spliced isoforms are mainly divided into two main classes: exon skipping and intron retention, where individual exons or introns are alternatively spliced or included in the maturation message.

To verify whether the HLA-G expression pattern of the above ccRCC patients constitutes a representative subset of the general profile of ccRCC patients, we compared our results with results obtained from a "renal carcinoma genome" (CAGEKID) study cohort including 100 ccRCC patients receiving treatment in four different european countries (czech republic, uk, romania and russia). The generated data constitutes a high quality resource enabling the detection of alternative splicing events with high accuracy (Scelo et al, 2014). Furthermore, we have evaluated in depth whether it is possible to investigate whether common factors of HLA-G (such as selecting an aligned pair of RNAseq data or a reference sequence) could potentially affect our analysis by using two different alignments BWA MEM and TopHat 2. The results confirm that data aligned to BWA MEM or TopHat2 yielded similar results (supplementary data). Further, read length counts at the individual level indicate a great similarity between the expression profiles of HLA-G transcripts found in our small scale ccRCC patient cohort and Cagekid cohort. These results are summarized in tables 2 and 3, and will be discussed more fully in the following sections.

2.4 intron retention events not described in expressed HLA-G transcripts

Intron retention is the most rare alternative splicing type in mammals and accounts for only about 3% of replacement transcripts [12 ]. So far, only the retention of intron 3 or intron 5 (previously referred to as intron 2 and intron 4 according to the IMGT/HLA nomenclature) has been reported in the HLA-G transcript literature. The transcripts retaining intron 3 encode HLA-G7[13] and those retaining intron 5 encode HLA-G5 and HLA-G6[7 ].

In our RNAseq analysis, the intron comprised by the exon was marked as retained. The results shown in FIG. 4 and summarized in Table 2 indicate that the read length retained was the most for introns 3 and 5. In addition, these data also support new overall findings from the retention of four additional introns: 1.4, 6 and 7. To verify the expression of intron-retained transcripts, we first looked for the presence of transcripts containing intron 1. To this end, we performed RT-PCR amplification using the strategy described in FIG. 5. First, a primer targeting intron 1 (Int1F) was used in combination with the reverse primer G257R of G257F [13 ]. Since the presence of introns may be due to contaminating endogenous genomic DNA, all samples were amplified in parallel with actin-specific primers located on two different exons. The expected size of the amplification of cDNA from mRNA is 320bp, while the expected size of genomic DNA is 560 bp. The results show that only a 320bp fragment was amplified in all samples, indicating the absence of genomic contamination (fig. 5B, left panel). In view of this result, we further amplified tumor samples using primers Int1F and G257R. An amplification band of the expected size (521bp) was obtained, consistent with the presence of intron 1 in HLA-G transcripts (FIG. 5B, right panel). This event has not been previously reported in the literature since HLA-G transcription initiation is associated only with exon 2 [26 ]. We did not detect a PCR amplified band of 649bp, which corresponds to the concomitant retention of intron 2. This is consistent with the results of RNAseq analysis, which indicates that intron 2 is rarely retained.

Table 2, read number of all observed HLA-G splicing events in ccRCC samples patients 1, 3, 4 and 5 were selected as representative samples with the aim of exploring the diversity of HLA-G isoforms. B00E4I3 and B00E4IS are the two samples with the highest HLA-G expression in CAGEKID (CAncer GEnome of the KIDney of the KIDney) [14 ].

Further analysis was performed to verify retention of intron 4 (fig. 5C). For this purpose, RT-PCR was performed using primer G257F in combination with a primer specifically targeting intron 4 (referred to as int 4R). Amplification with these primers yielded a 430bp DNA fragment (FIG. 5C, left panel), indicating the presence of intron 4 in HLA-G transcripts. The size of the amplified band is also consistent with the concomitant retention of intron 3. To further assess whether it is possible for the same transcript to retain multiple introns simultaneously, we performed RT-PCR amplification using primer int3F (whose sequence is complementary to the region of intron 3) in combination with primer int 4R. The result revealed a 380bp DNA fragment, which is expected to retain introns 3 and 4 in the same transcript (FIG. 5C, middle panel). In addition, amplification with the Int3F and Int5R primers yielded an amplification band of 725pb (fig. 5C, right panel). Notably, the size of this band corresponds to the retention of introns 3 and 5, excluding intron 4. These results clearly show that tumor samples may express transcripts that retain a single intron, while others retain transcripts of several different introns, which may not be identical. To the best of our knowledge, these events have not been described previously.

2.5 novel HLA-G transcripts having 5' -extension end

RNAseq data further revealed that some reads were aligned on either side of exon 1 (fig. 4). Transcripts derived from this region have not been previously reported. Indeed, the structure of this region remains a controversial issue, as the information contained in the Ensembl database suggests that HLA-G transcripts may start from this exon, which is located 5' of exon 1 as defined by the IMGT/HLA nomenclature (FIG. 1A). The presence or absence of this exon may result in major modifications, including promoter orientation, length of the 5' -untranslated region and transcription/translation initiation site. We assessed whether HLA-G transcripts could be initiated at this exon or even upstream by RT-PCR amplification. Two specific primers were designed. Primer Ex1F, the sequence of which is complementary to a region in exon 1(Ensembl database), and primer PrPr, the sequence of which is complementary to a region upstream of what is currently considered to be a promoter region (schematically represented in FIG. 1B). RT-PCR using these two upstream primers in combination with G526R produced two bands of the expected size: 690bp (for Ex1F-G526R) and 725bp (for PrPr-G526R), respectively (data not shown). To verify the specificity of these fragments, amplified DNA samples were sequenced and searched for nucleotide similarities in public databases using BLAST. The results demonstrated a high similarity to HLA-G, except that a 106bp fragment was deleted. Due to this deletion, the distance between the ATG located at the end of exon 1 and the ATG located at exon 2 was reduced from 118bp to 12bp (FIG. 6A). As a result, a deletion of 106bp made both ATGs satisfactory. This may now allow translation to start at the ATG located in the first exon and produce a protein with 5 other amino acids (MKTPR) in the 5' extension. Currently, the only translation initiation start site is due to the ATG in exon 2 (corresponding to exon 1 as defined by IMGT/HLA nomenclature). This transcript is also found in some but not all of the trophoblast cell samples tested. This suggests that factors regulating their expression remain to be elucidated.

Taken together, these results are consistent with the presence of a novel HLA-G transcript named HLA-G1L, with an extended 5' end, likely co-expressed with previously reported HLA-G isoforms in trophoblast and ccRCC tumor cells.

2.6 alternatively spliced exons potentially give rise to novel soluble HLA-G isoforms

Exon skipping is one of the major forms of alternative splicing, which results in multiple mRNA isoforms differing in the precise combination of their exon sequences. Here, we define an exon skipping event as the pairing between an exon-containing form and an exon-free form, which occurs on the same exon and has the same flanking intron. The same exon may be involved in multiple exon skipping events.

For HLA-G, only the phenomenon of skipping exon 4(HLA-G2), exon 5(HLA-G4) or both (HLA-G3) at the same time has been reported in the literature. In this study, read length matching to BWA mem revealed exon skipping that was never found. Table 2 reports the major jump events. We also confirmed these results by using TopHat 2.

The highest read coverage is consistent with skipping exon 7 alone, exon 7 containing the stop codon of the protein. However, since an additional in-frame stop codon was found at the beginning of exon 8, it is expected that the encoded protein lacking this exon will not be significantly modified. Most importantly, skipping exon 7 together with exon 6 encoding the transmembrane domain is highly relevant, as their deletion may result in isoforms lacking the transmembrane domain and cytoplasmic tail, and thus still constitute an unreported soluble protein.

When RT-PCR was performed using primer G963R, the sequence of which was complementary to the exon 6 region, in combination with the forward primer G257F (exon 3) or G256F (exon 4), no amplification product could be obtained. However, when primer G257F was used in combination with G526R, the expected 290bp amplified fragment was generated. These results, taken together, are consistent with HLA-G transcripts having exons 3 and 4 but lacking exon 6. Furthermore, when the samples of patient 1 were analyzed using these primers, amplified bands were obtained using the primer combination G526F-G963R, whereas no amplified band was detected using G257F-G963R, consistent with the expression of transcripts lacking exon 3.

2.7 alternatively spliced HLA-G isoforms lacking the α -1 Domain

Further analysis of the RNAseq data indicates that some reads may start at exon 4, as determined by quantifying the original count of reads within 20pb upstream of the exon receptor site.A predicted N-terminal truncated protein will lack the peptide signal and α domain.

Notably, the expression of these isoforms now provides a hypothesis for the differences in immunostaining patterns produced in certain tumor samples after labeling with 4H84 and antibodies raised against soluble isoforms, which was not previously explained by extensive knowledge of the structure of HLA-G isoforms.

Table 3, percentage of transcripts observed for each splicing event the percentage of the total sample is the percentage of the sample representing that event. The last two columns are the same indices calculated for 77 CAGEKID samples expressing HLA-G.

B.Tumor promotion of HLA-G isoforms

1. Materials and methods

1.1 production of lentiviruses expressing HLA-G isoforms

The HLA-G1 and HLA-G1L isoforms were introduced into the plasmid pWPXL (10510bp), located between the BamH1(3499) and NdeI (4334) sites, just 3' of the EF-1 α promoter, which directs the expression of the two subtypes HLA-G.

For HLA-G1:

the 3438bp insert comprises the HLA-G1 cDNA starting with the sequence SEQ ID NO.93AGTGTGGTACTTT and ending 3' with the sequence SEQ ID NO. 94TGGAAGACATGAGAACTTTCCA. This fragment is followed by a "red" variant of GFP (jellyfish green fluorescent protein, Victoria), named Synechocystis, already under the control of the CMV promoter. Finally, a molecular barcode was introduced at the 3' end as an integration marker and used for in vivo metastasis monitoring (Grosselin et al, Stem Cells, 10: 2162-71, 2013).

For HLA-G1L:

the insert of 3279bp comprises the HLA-G1L cDNA starting with the sequence SEQ ID NO.95ATATAGTAACATAGTGT and ending 3' with the sequence SEQ ID NO. 94TGGAAGACATGAGAACTTTCCA. This fragment was followed by a "blue" (cyan) variant of GFP, ECFP, which had bimodal excitation and emission spectra at 433/445nm and 475/503nm, making it a fluorescent dye with gloss and improved photostability. The ECFP is placed under the control of the CMV promoter. Finally, a molecular barcode was introduced at the 3' end as an integration marker and used for in vivo metastasis monitoring (Grosselin et al, Stem Cells, 10: 2162-71, 2013).

These two plasmids were used to produce lentiviral WPXL Δ U3 SIN, enveloped VSV-G, OGM group II, class 2 at 1.20E +08TU (transduction units)/ml.

2. Results

Each lentivirus contains a different HLA-G isoform. Lentiviruses were transduced into a well-characterized renal cell carcinoma clear cell line (RCC7 cells) that did not express HLA-G. Two independent transductions were performed for each isoform to improve the reliability and robustness of the results (robustness).

Each transduced RCC7 cell line was injected subcutaneously into 5 NSG mice under each condition. Untransduced RCC7 cells were used as controls.

Tumor/metastatic growth was assessed periodically after intradermal injection of cells.

At sacrifice of mice, tumor metastases and different tissues were removed for immunohistochemistry and expression (RNA) analysis. Each isoform is associated with a barcode, which can ensure that the obtained tumor and metastasis are from injected cells.

As shown in figure 7, xenografted mice from RCC7 line cells bearing long HLA-G1L isoforms showed more significant tumor growth at J38, at least in part associated with more developed intratumoral and peritumoral neovascularization. No intratumoral necrosis repair was observed in these tumors, unlike tumors produced by the RCC7 line carrying the HLA-G1 isoform.

Similar experiments were performed with nude mice. FIG. 8A shows photographs of nude mice xenografted with RCC7 cells expressing GFP, HLA-G1 or HLA-G1L at day 25 post-injection. FIG. 8B shows photographs of nude mice xenografted with RCC7 cells grown on basement membrane matrix and expressing GFP, HLA-G1 or HLA-G1L at day 25 post-injection.

C.Pro-angiogenic effects of HLA-G isoforms

RCC7 cells expressing GFP, HLA-G1 or HLA-G1L were prepared as described above (point B).

The left ear of NSG mice was injected with control (GFP expressing RCC7 cells) and the right ear with HLA-G1 or HLA-G1L expressing RCC7 cells. Photographs were taken on day 8. The results are shown in FIG. 9 (control RCC7 cells vs. HLA-G1L) and FIG. 10 (control RCC7 cells vs. HLA-G1).

The results demonstrate the angiogenic effect of HLA-G1L expression, which was not reproduced by HLA-G1 expression.

Reference to the literature

1.Rouas-Freiss,N.,et al.,Direct evidence to support the role of HLA-Gin protecting the fetus from maternal uterine natural killer cytolysis.ProcNatl Acad Sci U S A,1997.94(21):p.11520-5.

2.Ibrahim,E.C.,et al.,Tumor-specific up-regulation of thenonclassical class I HLA-G antigen expression in renal carcinoma.Cancer Res,2001.61(18):p.6838-45.

3.Bukur,J.,et al.,Functional role of human leukocyte antigen-G up-regulation in renal cell carcinoma.Cancer Res,2003.63(14):p.4107-11.

4.Brugarolas,J.,Molecular genetics of clear-cell renal cellcarcinoma.J Clin Oncol,2014.32(18):p.1968-76.

5.Agaugue,S.,E.D.Carosella,and N.Rouas-Freiss,Role of HLA-G in tumorescape through expansion of myeloid-derived suppressor cells and cytokinicbalance in favor of Th2versus Th1/Th17.Blood,2011.117(26):p.7021-31.

6.Loumagne,L.,et al.,In vivo evidence that secretion of HLA-G byimmunogenic tumor cells allows their evasion from immunosurveillance.Int JCancer,2014.135(9):p.2107-17.

7.Fujii,T.,A.Ishitani,and D.E.Geraghty,A soluble form of the HLA-Gantigen is encoded by a messenger ribonucleic acid containing intron 4.JImmunol,1994.153(12):p.5516-24.

8.Moch,H.,et al.,The 2016WHO Classification of Tumours of the UrinarySystem and Male Genital Organs-Part A:Renal,Penile,and Testicular Tumours.EurUrol,2016.70(1):p.93-105.

9.Krishnan,B.and L.D.Truong,Renal epithelial neoplasms:the diagnosticimplications of electron microscopic study in 55cases.Hum Pathol,2002.33(1):p.68-79.

10.Paul,P.,et al.,HLA-G,-E,-F preworkshop:tools and protocols foranalysis of non-classical class I genes transcription and proteinexpression.Hum Immunol,2000.61(11):p.1177-95.

11.Woolard,J.,et al.,Molecular diversity of VEGF-A as a regulator ofits biological activity.Microcirculation,2009.16(7):p.572-92.

12.Wong,J.J.,et al.,Intron retention in mRNA:No longer nonsense:Knownand putative roles of intron retention in normal and diseasebiology.Bioessays,2016.38(1):p.41-9.

13.Paul,P.,et al.,Identification of HLA-G7 as a new splice variant ofthe HLA-G mRNA and expression of soluble HLA-G5,-G6,and-G7 transcripts inhuman transfected cells.Hum Immunol,2000.61(11):p.1138-49.

14.Scelo,G.,et al.,Variation in genomic landscape of clear cell renalcell carcinoma across Europe.Nat Commun,2014.5:p.5135.

15.Nayar,R.,E.Bourtsos,and D.V.DeFrias,Hyaline globules in renal cellcarcinoma and hepatocellular carcinoma.A clue or a diagnostic pitfall onfine-needle aspiration？Am J Clin Pathol,2000.114(4):p.576-82.

16.Gerlinger,M.,et al.,Genomic architecture and evolution of clearcell renal cell carcinomas defined by multiregion sequencing.Nat Genet,2014.46(3):p.225-33.

17.Carosella,E.D.,et al.,HLA-G and HLA-E:fundamental andpathophysiological aspects.Immunol Today,2000.21(11):p.532-4.

18.Hoare,H.L.,et al.,Subtle changes in peptide conformationprofoundly affect recognition of the non-classical MHC class I molecule HLA-Eby the CD94-NKG2 natural killer cell receptors.J Mol Biol,2008.377(5):p.1297-303.

19.Kraemer,T.,et al.,HLA-E:Presentation of a Broader PeptideRepertoire Impacts the Cellular Immune Response-Implications on HSCTOutcome.Stem Cells Int,2015.2015:p.346714.

20.Li,H.and R.Durbin,Fast and accurate short read alignment withBurrows-Wheeler transform.Bioinformatics,2009.25(14):p.1754-60.

21.Li,H.,et al.,The Sequence Alignment/Map format andSAMtools.Bioinformatics,2009.25(16):p.2078-9.

22.Robinson,J.T.,et al.,Integrative genomics viewer.Nat Biotechnol,2011.29(1):p.24-6.

23.Morandi,F.,et al.,Recent Advances in Our Understanding of HLA-GBiology:Lessons from a Wide Spectrum of Human Diseases；Journal of ImmunologyResearch,vol.2016,Article ID 4326495,2016.

24.Kim,D.,Pertea,G.,Trapnell,C.,Pimentel,H.,Kelley,R.,and Salzberg,S.L.(2013).TopHat2:accurate alignment of transcriptomes in the presence ofinsertions,deletions and gene fusions.Genome biology 14,R36.

25.Wang,E.T.,Sandberg,R.,Luo,S.,Khrebtukova,I.,Zhang,L.,Mayr,C.,Kingsmore,S.F.,Schroth,G.P.,and Burge,C.B.(2008).Alternative isoformregulation in human tissue transcriptomes.Nature 456,470-476.

26.Geraghty,D.E.,Koller,B.H.,and Orr,H.T.(1987).A human majorhistocompatibility complex class I gene that encodes a protein with ashortened cytoplasmic segment.Proceedings of the National Academy of Sciencesof the United States of America 84,9145-9149.

27.Tatusova et al,"Blast 2 sequences-a new tool for comparing proteinand nucleotide sequences",FEMS Microbiol,1999,Lett.174:247-250.

28.Fons,P.,Chabot,S.,Cartwright,J.E.,Lenfant,F.,L'Faqihi,F.,Giustiniani,J.,Herault,J.,Gueguen,G.,Bono,F.,Savi,P.,Aguerre-Girr,M.,Fournel,S.,Malecaze,F.,Bensussan,A.,J.,&Le Bouteiller,P.(2006).Soluble HLA-G1inhibits angiogenesis through an apoptotic pathway and by direct binding toCD160receptor expressed by endothelial cells.Blood,108(8),2608-2615.

29.Tronik-Le Roux D,Renard J,Vérine J,et al.Novel landscape of HLA-Gisoforms expressed in clear cell renal cell carcinoma patients.MolecularOncology.

Sequence listing

<110> committee on atomic and alternative energy

French public standing assistance hospital

<120> HLA-G transcripts and isoforms and their uses

<130>F0263S00652/PCT

<150>EP17305986.6

<151>2017-07-24

<160>95

<170>PatentIn version 3.5

<210>1

<211>5

<212>PRT

<213> Intelligent people

<400>1

Met Lys Thr Pro Arg

1 5

<210>2

<211>24

<212>PRT

<213> Intelligent people

<400>2

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala

20

<210>3

<211>90

<212>PRT

<213> Intelligent people

<400>3

Gly Ser His Ser Met Arg Tyr Phe Ser Ala Ala Val Ser Arg Pro Gly

1 5 10 15

Arg Gly Glu Pro Arg Phe Ile Ala Met Gly Tyr Val Asp Asp Thr Gln

20 25 30

Phe Val Arg Phe Asp Ser Asp Ser Ala Cys Pro Arg Met Glu Pro Arg

35 40 45

Ala Pro Trp Val Glu Gln Glu Gly Pro Glu Tyr Trp Glu Glu Glu Thr

50 55 60

Arg Asn Thr Lys Ala His Ala Gln Thr Asp Arg Met Asn Leu Gln Thr

65 70 75 80

Leu Arg Gly Tyr Tyr Asn Gln Ser Glu Ala

85 90

<210>4

<211>92

<212>PRT

<213> Intelligent people

<400>4

Ser Ser His Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly Ser Asp

1 5 10 15

Gly Arg Leu Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly Lys Asp

20 2530

Tyr Leu Ala Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr

35 40 45

Ala Ala Gln Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val Ala Glu

50 55 60

Gln Arg Arg Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu His Arg

65 70 75 80

Tyr Leu Glu Asn Gly Lys Glu Met Leu Gln Arg Ala

85 90

<210>5

<211>92

<212>PRT

<213> Intelligent people

<400>5

Asp Pro Pro Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu

1 5 10 15

Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile

20 25 30

Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu

35 40 45

Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala

50 55 60

Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln

6570 75 80

His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp

85 90

<210>6

<211>40

<212>PRT

<213> Intelligent people

<400>6

Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly Ile Val Ala Gly

1 5 10 15

Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala Val Ala Ala Val

20 25 30

Leu Trp Arg Lys Lys Ser Ser Asp

35 40

<210>7

<211>343

<212>PRT

<213> Intelligent people

<400>7

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Ser Met Arg Tyr Phe Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu

35 40 45

Pro Arg Phe Ile Ala Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg

50 55 60

Phe Asp Ser Asp Ser Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp

65 70 75 80

Val Glu Gln Glu Gly Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr

85 90 95

Lys Ala His Ala Gln Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly

100 105 110

Tyr Tyr Asn Gln Ser Glu Ala Ser Ser His Thr Leu Gln Trp Met Ile

115 120 125

Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu Gln

130 135 140

Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu Arg

145 150 155 160

Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys Cys

165 170 175

Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly Thr

180 185 190

Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu Met Leu

195 200 205

Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro Val Phe

210 215 220

Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala

225 230 235 240

Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp

245 250 255

Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys

260 265 270

Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys

275 280 285

His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Lys

290 295 300

Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly Ile Val Ala Gly Leu

305 310 315 320

Val Val Leu Ala Ala Val Val Thr Gly Ala Ala Val Ala Ala Val Leu

325 330 335

Trp Arg Lys Lys Ser Ser Asp

340

<210>8

<211>304

<212>PRT

<213> Intelligent people

<400>8

Met Lys Thr Pro Arg Met Val Val Met Ala Pro ArgThr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Ser Met Arg Tyr Phe Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu

35 40 45

Pro Arg Phe Ile Ala Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg

50 55 60

Phe Asp Ser Asp Ser Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp

65 70 75 80

Val Glu Gln Glu Gly Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr

85 90 95

Lys Ala His Ala Gln Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly

100 105 110

Tyr Tyr Asn Gln Ser Glu Ala Ser Ser His Thr Leu Gln Trp Met Ile

115 120 125

Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu Gln

130 135 140

Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu Arg

145 150 155 160

Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys Cys

165 170 175

Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly Thr

180 185 190

Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu Met Leu

195 200 205

Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro Val Phe

210 215 220

Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala

225 230 235 240

Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp

245 250 255

Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys

260 265 270

Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys

275 280 285

His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Met

290 295 300

<210>9

<211>253

<212>PRT

<213> Intelligent people

<400>9

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu

35 40 45

Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala

50 55 60

Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln

65 70 75 80

Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg

85 90 95

Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu

100 105 110

Asn Gly Lys Glu Met Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val

115 120 125

Thr His His Pro Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala

130 135 140

Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly

145 150 155 160

Glu AspGln Thr Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly

165 170 175

Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu

180 185 190

Glu Gln Arg Tyr Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro

195 200 205

Leu Met Leu Arg Trp Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met

210 215 220

Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala

225 230 235 240

Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp

245 250

<210>10

<211>214

<212>PRT

<213> Intelligent people

<400>10

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu

35 40 45

Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala

50 55 60

Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln

65 70 75 80

Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg

85 90 95

Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu

100 105 110

Asn Gly Lys Glu Met Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val

115 120 125

Thr His His Pro Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala

130 135 140

Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly

145 150 155 160

Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly

165 170 175

Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu

180 185 190

Glu Gln Arg Tyr Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro

195 200 205

Leu Met Leu Arg Trp Met

210

<210>11

<211>161

<212>PRT

<213> Intelligent people

<400>11

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu

35 40 45

Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala

50 55 60

Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln

65 70 75 80

Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg

85 90 95

Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu

100 105 110

Asn Gly Lys Glu Met Leu Gln Arg Ala Glu Gln Ser Ser Leu Pro Thr

115 120 125

Ile Pro Ile Met Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val

130 135 140

Val Thr Gly Ala Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser

145 150 155 160

Asp

<210>12

<211>122

<212>PRT

<213> Intelligent people

<400>12

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu

35 40 45

Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala

50 55 60

Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln

65 70 75 80

Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg

85 90 95

Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu

100 105 110

Asn Gly Lys Glu Met Leu Gln Arg Ala Val

115 120

<210>13

<211>161

<212>PRT

<213> Intelligent people

<400>13

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Asp Pro Pro

20 25 30

Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr Leu

35 40 45

Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp

50 55 60

Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr

65 70 75 80

Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val

85 90 95

Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu Gly

100 105 110

Leu Pro Glu Pro Leu Met Leu Arg TrpLys Gln Ser Ser Leu Pro Thr

115 120 125

Ile Pro Ile Met Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val

130 135 140

Val Thr Gly Ala Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser

145 150 155 160

Asp

<210>14

<211>122

<212>PRT

<213> Intelligent people

<400>14

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Asp Pro Pro

20 25 30

Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr Leu

35 40 45

Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp

50 55 60

Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr

65 70 75 80

Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val

85 90 95

Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu Gly

100 105 110

Leu Pro Glu Pro Leu Met Leu Arg Trp Met

115 120

<210>15

<211>159

<212>PRT

<213> Intelligent people

<400>15

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Ser Met Arg Tyr Phe Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu

35 40 45

Pro Arg Phe Ile Ala Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg

50 55 60

Phe Asp Ser Asp Ser Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp

65 70 75 80

Val Glu Gln Glu Gly Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr

85 90 95

Lys Ala His Ala Gln Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly

100 105 110

Tyr Tyr Asn Gln Ser Glu Ala Lys Gln Ser Ser Leu Pro Thr Ile Pro

115 120 125

Ile Met Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr

130 135 140

Gly Ala Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp

145 150 155

<210>16

<211>120

<212>PRT

<213> Intelligent people

<400>16

Met Lys Thr Pro Arg Met Val Val Met Ala Pro Arg Thr Leu Phe Leu

1 5 10 15

Leu Leu Ser Gly Ala Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His

20 25 30

Ser Met Arg Tyr Phe Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu

35 40 45

Pro Arg Phe Ile Ala Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg

50 55 60

Phe Asp Ser Asp Ser Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp

65 70 75 80

Val Glu Gln Glu Gly Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr

85 90 95

Lys Ala His Ala Gln Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly

100 105 110

Tyr Tyr Asn Gln Ser Glu Ala Met

115 120

<210>17

<211>137

<212>PRT

<213> Intelligent people

<400>17

Met Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro

1 5 10 15

Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr

20 25 30

Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr

35 40 45

Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe

50 55 60

Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr

65 70 75 80

Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg

85 90 95

Trp Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly Ile Val Ala

100 105 110

Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala Val Ala Ala

115 120 125

Val Leu Trp Arg Lys Lys Ser Ser Asp

130 135

<210>18

<211>98

<212>PRT

<213> Intelligent people

<400>18

Met Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro

1 5 10 15

Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr

20 25 30

Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr

35 40 45

Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe

50 55 60

Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr

65 70 75 80

Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg

85 90 95

Trp Met

<210>19

<211>217

<212>PRT

<213> Intelligent people

<400>19

Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr

1 5 10 15

Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp

20 25 30

Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg

35 40 45

Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu

50 55 60

Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu

65 70 75 80

Met Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro

85 90 95

Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr

100 105 110

Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr

115 120 125

Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe

130 135 140

Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr

145 150 155 160

Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg

165 170 175

Trp Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly Ile Val Ala

180 185 190

Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala Val Ala Ala

195 200 205

Val Leu Trp Arg Lys Lys Ser Ser Asp

210 215

<210>20

<211>178

<212>PRT

<213> Intelligent people

<400>20

Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr

1 5 10 15

Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp

20 25 30

Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg

35 40 45

Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu

50 55 60

Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu

65 70 75 80

Met Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro

85 90 95

Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr

100 105 110

Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr

115 120 125

Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe

130 135 140

Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr

145 150 155 160

Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg

165 170 175

Trp Met

<210>21

<211>299

<212>PRT

<213> Intelligent people

<400>21

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 510 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Ser Met Arg Tyr Phe

20 25 30

Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu Pro Arg Phe Ile Ala

35 40 45

Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ser

50 55 60

Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp Val Glu Gln Glu Gly

65 70 75 80

Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr Lys Ala His Ala Gln

85 90 95

Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly Tyr Tyr Asn Gln Ser

100 105 110

Glu Ala Ser Ser His Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly

115 120 125

Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly

130 135 140

Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala

145 150 155 160

Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val

165 170175

Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu

180 185 190

His Arg Tyr Leu Glu Asn Gly Lys Glu Met Leu Gln Arg Ala Asp Pro

195 200 205

Pro Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr

210 215 220

Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr

225 230 235 240

Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu

245 250 255

Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val

260 265 270

Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu

275 280 285

Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Met

290 295

<210>22

<211>248

<212>PRT

<213> Intelligent people

<400>22

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

15 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Thr Leu Gln Trp Met

20 25 30

Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu

35 40 45

Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu

50 55 60

Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys

65 70 75 80

Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly

85 90 95

Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu Met

100 105 110

Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro Val

115 120 125

Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro

130 135 140

Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln

145 150 155 160

Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln

165170 175

Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr

180 185 190

Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp

195 200 205

Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly Ile Val Ala Gly

210 215 220

Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala Val Ala Ala Val

225 230 235 240

Leu Trp Arg Lys Lys Ser Ser Asp

245

<210>23

<211>209

<212>PRT

<213> Intelligent people

<400>23

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Thr Leu Gln Trp Met

20 25 30

Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu

35 40 45

Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu

5055 60

Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys

65 70 75 80

Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly

85 90 95

Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu Met

100 105 110

Leu Gln Arg Ala Asp Pro Pro Lys Thr His Val Thr His His Pro Val

115 120 125

Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro

130 135 140

Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln

145 150 155 160

Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln

165 170 175

Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr

180 185 190

Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp

195 200 205

Met

<210>24

<211>156

<212>PRT

<213> Intelligent people

<400>24

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Thr Leu Gln Trp Met

20 25 30

Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu

35 40 45

Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu

50 55 60

Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys

65 70 75 80

Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly

85 90 95

Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu Met

100 105 110

Leu Gln Arg Ala Glu Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly

115 120 125

Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala

130 135 140

Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp

145 150 155

<210>25

<211>117

<212>PRT

<213> Intelligent people

<400>25

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Thr Leu Gln Trp Met

20 25 30

Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu

35 40 45

Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp Leu

50 55 60

Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys

65 70 75 80

Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly

85 90 95

Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu Met

100 105 110

Leu Gln Arg Ala Val

115

<210>26

<211>156

<212>PRT

<213> Intelligent people

<400>26

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Asp Pro Pro Lys Thr His Val Thr

20 25 30

His His Pro Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu

35 40 45

Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu

50 55 60

Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp

65 70 75 80

Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu

85 90 95

Gln Arg Tyr Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu

100 105 110

Met Leu Arg Trp Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly

115 120 125

Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala

130 135 140

Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp

145 150 155

<210>27

<211>117

<212>PRT

<213> Intelligent people

<400>27

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Asp Pro Pro Lys Thr His Val Thr

20 25 30

His His Pro Val Phe Asp Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu

35 40 45

Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu

50 55 60

Asp Gln Thr Gln Asp Val Glu Leu Val Glu Thr Arg Pro Ala Gly Asp

65 70 75 80

Gly Thr Phe Gln Lys Trp Ala Ala Val Val Val Pro Ser Gly Glu Glu

85 90 95

Gln Arg Tyr Thr Cys His Val Gln His Glu Gly Leu Pro Glu Pro Leu

100 105 110

Met Leu Arg Trp Met

115

<210>28

<211>115

<212>PRT

<213> Intelligent people

<400>28

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Ser Met Arg Tyr Phe

20 25 30

Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu Pro Arg Phe Ile Ala

35 40 45

Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ser

50 55 60

Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp Val Glu Gln Glu Gly

65 70 75 80

Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr Lys Ala His Ala Gln

85 90 95

Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly Tyr Tyr Asn Gln Ser

100 105 110

Glu Ala Met

115

<210>29

<211>125

<212>PRT

<213> Intelligent people

<400>29

Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr

1 5 10 15

Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp

20 25 30

Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg

35 40 45

Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu

50 55 60

Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu

65 70 75 80

Met Leu Gln Arg Ala Glu Gln Ser Ser Leu Pro Thr Ile Pro Ile Met

85 90 95

Gly Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala

100 105 110

Ala Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp

115 120 125

<210>30

<211>86

<212>PRT

<213> Intelligent people

<400>30

Met Ile Gly Cys Asp Leu Gly Ser Asp Gly Arg Leu Leu Arg Gly Tyr

15 10 15

Glu Gln Tyr Ala Tyr Asp Gly Lys Asp Tyr Leu Ala Leu Asn Glu Asp

20 25 30

Leu Arg Ser Trp Thr Ala Ala Asp Thr Ala Ala Gln Ile Ser Lys Arg

35 40 45

Lys Cys Glu Ala Ala Asn Val Ala Glu Gln Arg Arg Ala Tyr Leu Glu

50 55 60

Gly Thr Cys Val Glu Trp Leu His Arg Tyr Leu Glu Asn Gly Lys Glu

65 70 75 80

Met Leu Gln Arg Ala Val

85

<210>31

<211>44

<212>PRT

<213> Intelligent people

<400>31

Met Leu Arg Trp Lys Gln Ser Ser Leu Pro Thr Ile Pro Ile Met Gly

1 5 10 15

Ile Val Ala Gly Leu Val Val Leu Ala Ala Val Val Thr Gly Ala Ala

20 25 30

Val Ala Ala Val Leu Trp Arg Lys Lys Ser Ser Asp

35 40

<210>32

<211>40

<212>DNA

<213> Intelligent people

<400>32

ctgaccgaga cctgggcggg ttctcacacc ctccagtgga 40

<210>33

<211>33

<212>DNA

<213> Intelligent people

<400>33

ccgagacctg ggcggacccc cccaagacac acg 33

<210>34

<211>32

<212>DNA

<213> Intelligent people

<400>34

tcatgctgag atggatgtga aacagctgcc ct 32

<210>35

<211>9

<212>PRT

<213> Intelligent people

<400>35

Trp Ala Gly Ser His Thr Leu Gln Trp

1 5

<210>36

<211>10

<212>PRT

<213> Intelligent people

<400>36

Glu Thr Trp Ala Asp Pro Pro Lys Thr His

1 5 10

<210>37

<211>5

<212>PRT

<213> Intelligent people

<400>37

Met Leu Arg Trp Met

1 5

<210>38

<211>1490

<212>DNA

<213> Intelligent people

<400>38

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gctcccactc catgaggtat 180

ttcagcgccg ccgtgtcccg gcccggccgc ggggagcccc gcttcatcgc catgggctac 240

gtggacgaca cgcagttcgt gcggttcgac agcgactcgg cgtgtccgag gatggagccg 300

cgggcgccgt gggtggagca ggaggggccg gagtattggg aagaggagac acggaacacc 360

aaggcccacg cacagactga cagaatgaac ctgcagaccc tgcgcggcta ctacaaccag 420

agcgaggcca gttctcacac cctccagtgg atgattggct gcgacctggg gtccgacgga 480

cgcctcctcc gcgggtatga acagtatgcc tacgatggca aggattacct cgccctgaac 540

gaggacctgc gctcctggac cgcagcggac actgcggctc agatctccaa gcgcaagtgt 600

gaggcggcca atgtggctga acaaaggaga gcctacctgg agggcacgtg cgtggagtgg 660

ctccacagat acctggagaa cgggaaggag atgctgcagc gcgcggaccc ccccaagaca 720

cacgtgaccc accaccctgt ctttgactat gaggccaccc tgaggtgctg ggccctgggc 780

ttctaccctgcggagatcat actgacctgg cagcgggatg gggaggacca gacccaggac 840

gtggagctcg tggagaccag gcctgcaggg gatggaacct tccagaagtg ggcagctgtg 900

gtggtgcctt ctggagagga gcagagatac acgtgccatg tgcagcatga ggggctgccg 960

gagcccctca tgctgagatg gaagcagtct tccctgccca ccatccccat catgggtatc 1020

gttgctggcc tggttgtcct tgcagctgta gtcactggag ctgcggtcgc tgctgtgctg 1080

tggagaaaga agagctcaga ttgaaaagga gggagctact ctcaggctgc aatgtgaaac 1140

agctgccctg tgtgggactg agtggcaagt ccctttgtga cttcaagaac cctgactcct 1200

ctttgtgcag agaccagccc acccctgtgc ccaccatgac cctcttcctc atgctgaact 1260

gcattccttc cccaatcacc tttcctgttc cagaaaaggg gctgggatgt ctccgtctct 1320

gtctcaaatt tgtggtccac tgagctataa cttacttctg tattaaaatt agaatctgag 1380

tataaattta ctttttcaaa ttatttccaa gagagattga tgggttaatt aaaggagaag 1440

attcctgaaa tttgagagac aaaataaatg gaagacatga gaactttcca 1490

<210>39

<211>1340

<212>DNA

<213> Intelligent people

<400>39

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gctcccactc catgaggtat 180

ttcagcgccg ccgtgtcccg gcccggccgc ggggagcccc gcttcatcgc catgggctac 240

gtggacgaca cgcagttcgt gcggttcgac agcgactcgg cgtgtccgag gatggagccg 300

cgggcgccgt gggtggagca ggaggggccg gagtattggg aagaggagac acggaacacc 360

aaggcccacg cacagactga cagaatgaac ctgcagaccc tgcgcggcta ctacaaccag 420

agcgaggcca gttctcacac cctccagtgg atgattggct gcgacctggg gtccgacgga 480

cgcctcctcc gcgggtatga acagtatgcc tacgatggca aggattacct cgccctgaac 540

gaggacctgc gctcctggac cgcagcggac actgcggctc agatctccaa gcgcaagtgt 600

gaggcggcca atgtggctga acaaaggaga gcctacctgg agggcacgtg cgtggagtgg 660

ctccacagat acctggagaa cgggaaggag atgctgcagc gcgcggaccc ccccaagaca 720

cacgtgaccc accaccctgt ctttgactat gaggccaccc tgaggtgctg ggccctgggc 780

ttctaccctg cggagatcat actgacctgg cagcgggatg gggaggacca gacccaggac 840

gtggagctcg tggagaccag gcctgcaggg gatggaacct tccagaagtg ggcagctgtg 900

gtggtgcctt ctggagagga gcagagatac acgtgccatg tgcagcatga ggggctgccg 960

gagcccctca tgctgagatg gatgtgaaac agctgccctg tgtgggactg agtggcaagt 1020

ccctttgtga cttcaagaac cctgactcct ctttgtgcag agaccagccc acccctgtgc 1080

ccaccatgac cctcttcctc atgctgaact gcattccttc cccaatcacc tttcctgttc 1140

cagaaaaggg gctgggatgt ctccgtctct gtctcaaatt tgtggtccac tgagctataa 1200

cttacttctg tattaaaatt agaatctgag tataaattta ctttttcaaa ttatttccaa 1260

gagagattga tgggttaatt aaaggagaag attcctgaaa tttgagagac aaaataaatg 1320

gaagacatga gaactttcca 1340

<210>40

<211>1220

<212>DNA

<213> Intelligent people

<400>40

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gttctcacac cctccagtgg 180

atgattggct gcgacctggg gtccgacgga cgcctcctcc gcgggtatga acagtatgcc 240

tacgatggca aggattacct cgccctgaac gaggacctgc gctcctggac cgcagcggac 300

actgcggctc agatctccaa gcgcaagtgt gaggcggcca atgtggctga acaaaggaga 360

gcctacctgg agggcacgtg cgtggagtgg ctccacagat acctggagaa cgggaaggag 420

atgctgcagc gcgcggaccc ccccaagaca cacgtgaccc accaccctgt ctttgactat 480

gaggccaccc tgaggtgctg ggccctgggc ttctaccctg cggagatcat actgacctgg 540

cagcgggatg gggaggacca gacccaggac gtggagctcg tggagaccag gcctgcaggg 600

gatggaacct tccagaagtg ggcagctgtg gtggtgcctt ctggagagga gcagagatac 660

acgtgccatg tgcagcatga ggggctgccg gagcccctca tgctgagatg gaagcagtct 720

tccctgccca ccatccccat catgggtatc gttgctggcc tggttgtcct tgcagctgta 780

gtcactggag ctgcggtcgc tgctgtgctg tggagaaaga agagctcaga ttgaaaagga 840

gggagctact ctcaggctgc aatgtgaaac agctgccctg tgtgggactg agtggcaagt 900

ccctttgtga cttcaagaac cctgactcct ctttgtgcag agaccagccc acccctgtgc 960

ccaccatgac cctcttcctc atgctgaact gcattccttc cccaatcacc tttcctgttc 1020

cagaaaaggg gctgggatgt ctccgtctct gtctcaaatt tgtggtccac tgagctataa 1080

cttacttctg tattaaaatt agaatctgag tataaattta ctttttcaaa ttatttccaa 1140

gagagattga tgggttaatt aaaggagaag attcctgaaa tttgagagac aaaataaatg 1200

gaagacatga gaactttcca 1220

<210>41

<211>1070

<212>DNA

<213> Intelligent people

<400>41

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gttctcacac cctccagtgg 180

atgattggct gcgacctggg gtccgacgga cgcctcctcc gcgggtatga acagtatgcc 240

tacgatggca aggattacct cgccctgaac gaggacctgc gctcctggac cgcagcggac 300

actgcggctc agatctccaa gcgcaagtgt gaggcggcca atgtggctga acaaaggaga 360

gcctacctgg agggcacgtg cgtggagtgg ctccacagat acctggagaa cgggaaggag 420

atgctgcagc gcgcggaccc ccccaagaca cacgtgaccc accaccctgt ctttgactat 480

gaggccaccc tgaggtgctg ggccctgggc ttctaccctg cggagatcat actgacctgg 540

cagcgggatg gggaggacca gacccaggac gtggagctcg tggagaccag gcctgcaggg 600

gatggaacct tccagaagtg ggcagctgtg gtggtgcctt ctggagagga gcagagatac 660

acgtgccatg tgcagcatga ggggctgccg gagcccctca tgctgagatg gatgtgaaac 720

agctgccctg tgtgggactg agtggcaagt ccctttgtga cttcaagaac cctgactcct 780

ctttgtgcag agaccagccc acccctgtgc ccaccatgac cctcttcctc atgctgaact 840

gcattccttc cccaatcacc tttcctgttc cagaaaaggg gctgggatgt ctccgtctct 900

gtctcaaatt tgtggtccac tgagctataa cttacttctg tattaaaatt agaatctgag 960

tataaattta ctttttcaaa ttatttccaa gagagattga tgggttaatt aaaggagaag 1020

attcctgaaa tttgagagac aaaataaatg gaagacatga gaactttcca 1070

<210>42

<211>944

<212>DNA

<213> Intelligent people

<400>42

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gttctcacac cctccagtgg 180

atgattggct gcgacctggg gtccgacgga cgcctcctcc gcgggtatga acagtatgcc 240

tacgatggca aggattacct cgccctgaac gaggacctgc gctcctggac cgcagcggac 300

actgcggctc agatctccaa gcgcaagtgt gaggcggcca atgtggctga acaaaggaga 360

gcctacctgg agggcacgtg cgtggagtgg ctccacagat acctggagaa cgggaaggag 420

atgctgcagc gcgcggagca gtcttccctg cccaccatcc ccatcatggg tatcgttgct 480

ggcctggttg tccttgcagc tgtagtcact ggagctgcgg tcgctgctgt gctgtggaga 540

aagaagagct cagattgaaa aggagggagc tactctcagg ctgcaatgtg aaacagctgc 600

cctgtgtggg actgagtggc aagtcccttt gtgacttcaa gaaccctgac tcctctttgt 660

gcagagacca gcccacccct gtgcccacca tgaccctctt cctcatgctg aactgcattc 720

cttccccaat cacctttcct gttccagaaa aggggctggg atgtctccgt ctctgtctca 780

aatttgtggt ccactgagct ataacttact tctgtattaa aattagaatc tgagtataaa 840

tttacttttt caaattattt ccaagagaga ttgatgggtt aattaaagga gaagattcct 900

gaaatttgag agacaaaata aatggaagac atgagaactt tcca 944

<210>43

<211>794

<212>DNA

<213> Intelligent people

<400>43

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gttctcacac cctccagtgg 180

atgattggct gcgacctggg gtccgacgga cgcctcctcc gcgggtatga acagtatgcc 240

tacgatggca aggattacct cgccctgaac gaggacctgc gctcctggac cgcagcggac 300

actgcggctc agatctccaa gcgcaagtgt gaggcggcca atgtggctga acaaaggaga 360

gcctacctgg agggcacgtg cgtggagtgg ctccacagat acctggagaa cgggaaggag 420

atgctgcagc gcgcggtgtg aaacagctgc cctgtgtggg actgagtggc aagtcccttt 480

gtgacttcaa gaaccctgac tcctctttgt gcagagacca gcccacccct gtgcccacca 540

tgaccctctt cctcatgctg aactgcattc cttccccaat cacctttcct gttccagaaa 600

aggggctggg atgtctccgt ctctgtctca aatttgtggt ccactgagct ataacttact 660

tctgtattaa aattagaatc tgagtataaa tttacttttt caaattattt ccaagagaga 720

ttgatgggtt aattaaagga gaagattcct gaaatttgag agacaaaata aatggaagac 780

atgagaactt tcca 794

<210>44

<211>944

<212>DNA

<213> Intelligent people

<400>44

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg acccccccaa gacacacgtg 180

acccaccacc ctgtctttga ctatgaggcc accctgaggt gctgggccct gggcttctac 240

cctgcggaga tcatactgac ctggcagcgg gatggggagg accagaccca ggacgtggag 300

ctcgtggaga ccaggcctgc aggggatgga accttccaga agtgggcagc tgtggtggtg 360

ccttctggag aggagcagag atacacgtgc catgtgcagc atgaggggct gccggagccc 420

ctcatgctga gatggaagca gtcttccctg cccaccatcc ccatcatggg tatcgttgct 480

ggcctggttg tccttgcagc tgtagtcact ggagctgcgg tcgctgctgt gctgtggaga 540

aagaagagct cagattgaaa aggagggagc tactctcagg ctgcaatgtg aaacagctgc 600

cctgtgtggg actgagtggc aagtcccttt gtgacttcaa gaaccctgac tcctctttgt 660

gcagagacca gcccacccct gtgcccacca tgaccctctt cctcatgctg aactgcattc 720

cttccccaat cacctttcct gttccagaaa aggggctggg atgtctccgt ctctgtctca 780

aatttgtggt ccactgagct ataacttact tctgtattaa aattagaatc tgagtataaa 840

tttacttttt caaattattt ccaagagaga ttgatgggtt aattaaagga gaagattcct 900

gaaatttgag agacaaaata aatggaagac atgagaactt tcca 944

<210>45

<211>794

<212>DNA

<213> Intelligent people

<400>45

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg acccccccaa gacacacgtg 180

acccaccacc ctgtctttga ctatgaggcc accctgaggt gctgggccct gggcttctac 240

cctgcggaga tcatactgac ctggcagcgg gatggggagg accagaccca ggacgtggag 300

ctcgtggaga ccaggcctgc aggggatgga accttccaga agtgggcagc tgtggtggtg 360

ccttctggag aggagcagag atacacgtgc catgtgcagc atgaggggct gccggagccc 420

ctcatgctga gatggatgtg aaacagctgc cctgtgtggg actgagtggc aagtcccttt 480

gtgacttcaa gaaccctgac tcctctttgt gcagagacca gcccacccct gtgcccacca 540

tgaccctctt cctcatgctg aactgcattc cttccccaat cacctttcct gttccagaaa 600

aggggctggg atgtctccgt ctctgtctca aatttgtggt ccactgagct ataacttact 660

tctgtattaa aattagaatc tgagtataaa tttacttttt caaattattt ccaagagaga 720

ttgatgggtt aattaaagga gaagattcct gaaatttgag agacaaaata aatggaagac 780

atgagaactt tcca 794

<210>46

<211>938

<212>DNA

<213> Intelligent people

<400>46

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gctcccactc catgaggtat 180

ttcagcgccg ccgtgtcccg gcccggccgc ggggagcccc gcttcatcgc catgggctac 240

gtggacgaca cgcagttcgt gcggttcgac agcgactcgg cgtgtccgag gatggagccg 300

cgggcgccgt gggtggagca ggaggggccg gagtattggg aagaggagac acggaacacc 360

aaggcccacg cacagactga cagaatgaac ctgcagaccc tgcgcggcta ctacaaccag 420

agcgaggcca agcagtcttc cctgcccacc atccccatca tgggtatcgt tgctggcctg 480

gttgtccttg cagctgtagt cactggagct gcggtcgctg ctgtgctgtg gagaaagaag 540

agctcagatt gaaaaggagg gagctactct caggctgcaa tgtgaaacag ctgccctgtg 600

tgggactgag tggcaagtcc ctttgtgact tcaagaaccc tgactcctct ttgtgcagag 660

accagcccac ccctgtgccc accatgaccc tcttcctcat gctgaactgc attccttccc 720

caatcacctt tcctgttcca gaaaaggggc tgggatgtct ccgtctctgt ctcaaatttg 780

tggtccactg agctataact tacttctgta ttaaaattag aatctgagta taaatttact 840

ttttcaaatt atttccaaga gagattgatg ggttaattaa aggagaagat tcctgaaatt 900

tgagagacaa aataaatgga agacatgaga actttcca 938

<210>47

<211>788

<212>DNA

<213> Intelligent people

<400>47

atatagtaac atagtgtggt actttgtctt gaggagatgt cctggactca cacggaaact 60

tagggctacg gaatgaagac gccaaggatg gtggtcatgg cgccccgaac cctcttcctg 120

ctgctctcgg gggccctgac cctgaccgag acctgggcgg gctcccactc catgaggtat 180

ttcagcgccg ccgtgtcccg gcccggccgc ggggagcccc gcttcatcgc catgggctac 240

gtggacgaca cgcagttcgt gcggttcgac agcgactcgg cgtgtccgag gatggagccg 300

cgggcgccgt gggtggagca ggaggggccg gagtattggg aagaggagac acggaacacc 360

aaggcccacg cacagactga cagaatgaac ctgcagaccc tgcgcggcta ctacaaccag 420

agcgaggcca tgtgaaacag ctgccctgtg tgggactgag tggcaagtcc ctttgtgact 480

tcaagaaccc tgactcctct ttgtgcagag accagcccac ccctgtgccc accatgaccc 540

tcttcctcat gctgaactgc attccttccc caatcacctt tcctgttcca gaaaaggggc 600

tgggatgtct ccgtctctgt ctcaaatttg tggtccactg agctataact tacttctgta 660

ttaaaattag aatctgagta taaatttact ttttcaaatt atttccaaga gagattgatg 720

ggttaattaa aggagaagat tcctgaaatt tgagagacaa aataaatgga agacatgaga 780

actttcca 788

<210>48

<211>1427

<212>DNA

<213> Intelligent people

<400>48

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggct cccactccat gaggtatttc 120

agcgccgccg tgtcccggcc cggccgcggg gagccccgct tcatcgccat gggctacgtg 180

gacgacacgc agttcgtgcg gttcgacagc gactcggcgt gtccgaggat ggagccgcgg 240

gcgccgtggg tggagcagga ggggccggag tattgggaag aggagacacg gaacaccaag 300

gcccacgcac agactgacag aatgaacctg cagaccctgc gcggctacta caaccagagc 360

gaggccagtt ctcacaccct ccagtggatg attggctgcg acctggggtc cgacggacgc 420

ctcctccgcg ggtatgaaca gtatgcctac gatggcaagg attacctcgc cctgaacgag 480

gacctgcgct cctggaccgc agcggacact gcggctcaga tctccaagcg caagtgtgag 540

gcggccaatg tggctgaaca aaggagagcc tacctggagg gcacgtgcgt ggagtggctc 600

cacagatacc tggagaacgg gaaggagatg ctgcagcgcg cggacccccc caagacacac 660

gtgacccaccaccctgtctt tgactatgag gccaccctga ggtgctgggc cctgggcttc 720

taccctgcgg agatcatact gacctggcag cgggatgggg aggaccagac ccaggacgtg 780

gagctcgtgg agaccaggcc tgcaggggat ggaaccttcc agaagtgggc agctgtggtg 840

gtgccttctg gagaggagca gagatacacg tgccatgtgc agcatgaggg gctgccggag 900

cccctcatgc tgagatggaa gcagtcttcc ctgcccacca tccccatcat gggtatcgtt 960

gctggcctgg ttgtccttgc agctgtagtc actggagctg cggtcgctgc tgtgctgtgg 1020

agaaagaaga gctcagattg aaaaggaggg agctactctc aggctgcaat gtgaaacagc 1080

tgccctgtgt gggactgagt ggcaagtccc tttgtgactt caagaaccct gactcctctt 1140

tgtgcagaga ccagcccacc cctgtgccca ccatgaccct cttcctcatg ctgaactgca 1200

ttccttcccc aatcaccttt cctgttccag aaaaggggct gggatgtctc cgtctctgtc 1260

tcaaatttgt ggtccactga gctataactt acttctgtat taaaattaga atctgagtat 1320

aaatttactt tttcaaatta tttccaagag agattgatgg gttaattaaa ggagaagatt 1380

cctgaaattt gagagacaaa ataaatggaa gacatgagaa ctttcca 1427

<210>49

<211>1276

<212>DNA

<213> Intelligent people

<400>49

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggct cccactccat gaggtatttc 120

agcgccgccg tgtcccggcc cggccgcggg gagccccgct tcatcgccat gggctacgtg 180

gacgacacgc agttcgtgcg gttcgacagc gactcggcgt gtccgaggat ggagccgcgg 240

gcgccgtggg tggagcagga ggggccggag tattgggaag aggagacacg gaacaccaag 300

gcccacgcac agactgacag aatgaacctg cagaccctgc gcggctacta caaccagagc 360

gaggccagtt ctcacaccct ccagtggatg attggctgcg acctggggtc cgacggacgc 420

ctcctccgcg ggtatgaaca gtatgcctac gatggcaagg attacctcgc cctgaacgag 480

gacctgcgct cctggaccgc agcggacact gcggctcaga tctccaagcg caagtgtgag 540

gcggccaatg tggctgaaca aaggagagcc tacctggagg gcacgtgcgt ggagtggctc 600

cacagatacc tggagaacgg gaaggagatg ctgcagcgcg cggacccccc caagacacac 660

gtgacccacc accctgtctt tgactatgag gccaccctga ggtgctgggc cctgggcttc 720

taccctgcgg agatcatact gacctggcag cgggatggga ggaccagacc caggacgtgg 780

agctcgtgga gaccaggcct gcaggggatg gaaccttcca gaagtgggca gctgtggtgg 840

tgccttctgg agaggagcag agatacacgt gccatgtgca gcatgagggg ctgccggagc 900

ccctcatgct gagatggatg tgaaacagct gccctgtgtg ggactgagtg gcaagtccct 960

ttgtgacttc aagaaccctg actcctcttt gtgcagagac cagcccaccc ctgtgcccac 1020

catgaccctc ttcctcatgc tgaactgcat tccttcccca atcacctttc ctgttccaga 1080

aaaggggctg ggatgtctcc gtctctgtct caaatttgtg gtccactgag ctataactta 1140

cttctgtatt aaaattagaa tctgagtata aatttacttt ttcaaattat ttccaagaga 1200

gattgatggg ttaattaaag gagaagattc ctgaaatttg agagacaaaa taaatggaag 1260

acatgagaac tttcca 1276

<210>50

<211>1157

<212>DNA

<213> Intelligent people

<400>50

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggtt ctcacaccct ccagtggatg 120

attggctgcg acctggggtc cgacggacgc ctcctccgcg ggtatgaaca gtatgcctac 180

gatggcaagg attacctcgc cctgaacgag gacctgcgct cctggaccgc agcggacact 240

gcggctcaga tctccaagcg caagtgtgag gcggccaatg tggctgaaca aaggagagcc 300

tacctggagg gcacgtgcgt ggagtggctc cacagatacc tggagaacgg gaaggagatg 360

ctgcagcgcg cggacccccc caagacacac gtgacccacc accctgtctt tgactatgag 420

gccaccctga ggtgctgggc cctgggcttc taccctgcgg agatcatact gacctggcag 480

cgggatgggg aggaccagac ccaggacgtg gagctcgtgg agaccaggcc tgcaggggat 540

ggaaccttcc agaagtgggc agctgtggtg gtgccttctg gagaggagca gagatacacg 600

tgccatgtgc agcatgaggg gctgccggag cccctcatgc tgagatggaa gcagtcttcc 660

ctgcccacca tccccatcat gggtatcgtt gctggcctgg ttgtccttgc agctgtagtc 720

actggagctg cggtcgctgc tgtgctgtgg agaaagaaga gctcagattg aaaaggaggg 780

agctactctc aggctgcaat gtgaaacagc tgccctgtgt gggactgagt ggcaagtccc 840

tttgtgactt caagaaccct gactcctctt tgtgcagaga ccagcccacc cctgtgccca 900

ccatgaccct cttcctcatg ctgaactgca ttccttcccc aatcaccttt cctgttccag 960

aaaaggggct gggatgtctc cgtctctgtc tcaaatttgt ggtccactga gctataactt 1020

acttctgtat taaaattaga atctgagtat aaatttactt tttcaaatta tttccaagag 1080

agattgatgg gttaattaaa ggagaagatt cctgaaattt gagagacaaa ataaatggaa 1140

gacatgagaa ctttcca 1157

<210>51

<211>1040

<212>DNA

<213> Intelligent people

<400>51

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggtt ctcacaccct ccagtggatg 120

attggctgcg acctggggtc cgacggacgc ctcctccgcg ggtatgaaca gtatgcctac 180

gatggcaagg attacctcgc cctgaacgag gacctgcgct cctggaccgc agcggacact 240

gcggctcaga tctccaagcg caagtgtgag gcggccaatg tggctgaaca aaggagagcc 300

tacctggagg gcacgtgcgt ggagtggctc cacagatacc tggagaacgg gaaggagatg 360

ctgcagcgcg cggacccccc caagacacac gtgacccacc accctgtctt tgactatgag 420

gccaccctga ggtgctgggc cctgggcttc taccctgcgg agatcatact gacctggcag 480

cgggatgggg aggaccagac ccaggacgtg gagctcgtgg agaccaggcc tgcaggggat 540

ggaaccttcc agaagtgggc agctgtggtg gtgccttctg gagaggagca gagatacacg 600

tgccatgtgc agcatgaggg gctgccggag cccctcatgc tgagatggaa ttgaaaagga 660

gggagctact ctcaggctgc aatgtgaaac agctgccctg tgtgggactg agtggcaagt 720

ccctttgtga cttcaagaac cctgactcct ctttgtgcag agaccagccc acccctgtgc 780

ccaccatgac cctcttcctc atgctgaact gcattccttc cccaatcacc tttcctgttc 840

cagaaaaggg gctgggatgt ctccgtctct gtctcaaatt tgtggtccac tgagctataa 900

cttacttctg tattaaaatt agaatctgag tataaattta ctttttcaaa ttatttccaa 960

gagagattga tgggttaatt aaaggagaag attcctgaaa tttgagagac aaaataaatg 1020

gaagacatga gaactttcca 1040

<210>52

<211>881

<212>DNA

<213> Intelligent people

<400>52

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggtt ctcacaccct ccagtggatg 120

attggctgcg acctggggtc cgacggacgc ctcctccgcg ggtatgaaca gtatgcctac 180

gatggcaagg attacctcgc cctgaacgag gacctgcgct cctggaccgc agcggacact 240

gcggctcaga tctccaagcg caagtgtgag gcggccaatg tggctgaaca aaggagagcc 300

tacctggagg gcacgtgcgt ggagtggctc cacagatacc tggagaacgg gaaggagatg 360

ctgcagcgcg cggagcagtc ttccctgccc accatcccca tcatgggtat cgttgctggc 420

ctggttgtcc ttgcagctgt agtcactgga gctgcggtcg ctgctgtgct gtggagaaag 480

aagagctcag attgaaaagg agggagctac tctcaggctg caatgtgaaa cagctgccct 540

gtgtgggact gagtggcaag tccctttgtg acttcaagaa ccctgactcc tctttgtgca 600

gagaccagcc cacccctgtg cccaccatga ccctcttcct catgctgaac tgcattcctt 660

ccccaatcac ctttcctgtt ccagaaaagg ggctgggatg tctccgtctc tgtctcaaat 720

ttgtggtcca ctgagctata acttacttct gtattaaaat tagaatctga gtataaattt 780

actttttcaa attatttcca agagagattg atgggttaat taaaggagaa gattcctgaa 840

atttgagaga caaaataaat ggaagacatg agaactttcc a 881

<210>53

<211>731

<212>DNA

<213> Intelligent people

<400>53

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggtt ctcacaccct ccagtggatg 120

attggctgcg acctggggtc cgacggacgc ctcctccgcg ggtatgaaca gtatgcctac 180

gatggcaagg attacctcgc cctgaacgag gacctgcgct cctggaccgc agcggacact 240

gcggctcaga tctccaagcg caagtgtgag gcggccaatg tggctgaaca aaggagagcc 300

tacctggagg gcacgtgcgt ggagtggctc cacagatacc tggagaacgg gaaggagatg 360

ctgcagcgcg cggtgtgaaa cagctgccct gtgtgggact gagtggcaag tccctttgtg 420

acttcaagaa ccctgactcc tctttgtgca gagaccagcc cacccctgtg cccaccatga 480

ccctcttcct catgctgaac tgcattcctt ccccaatcac ctttcctgtt ccagaaaagg 540

ggctgggatg tctccgtctc tgtctcaaat ttgtggtcca ctgagctata acttacttct 600

gtattaaaat tagaatctga gtataaattt actttttcaa attatttcca agagagattg 660

atgggttaat taaaggagaa gattcctgaa atttgagaga caaaataaat ggaagacatg 720

agaactttcc a 731

<210>54

<211>881

<212>DNA

<213> Intelligent people

<400>54

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcggacc cccccaagac acacgtgacc 120

caccaccctg tctttgacta tgaggccacc ctgaggtgct gggccctggg cttctaccct 180

gcggagatca tactgacctg gcagcgggat ggggaggacc agacccagga cgtggagctc 240

gtggagacca ggcctgcagg ggatggaacc ttccagaagt gggcagctgt ggtggtgcct 300

tctggagagg agcagagata cacgtgccat gtgcagcatg aggggctgcc ggagcccctc 360

atgctgagat ggaagcagtc ttccctgccc accatcccca tcatgggtat cgttgctggc 420

ctggttgtcc ttgcagctgt agtcactgga gctgcggtcg ctgctgtgct gtggagaaag 480

aagagctcag attgaaaagg agggagctac tctcaggctg caatgtgaaa cagctgccct 540

gtgtgggact gagtggcaag tccctttgtg acttcaagaa ccctgactcc tctttgtgca 600

gagaccagcc cacccctgtg cccaccatga ccctcttcct catgctgaac tgcattcctt 660

ccccaatcac ctttcctgtt ccagaaaagg ggctgggatg tctccgtctc tgtctcaaat 720

ttgtggtcca ctgagctata acttacttct gtattaaaat tagaatctga gtataaattt 780

actttttcaa attatttcca agagagattg atgggttaat taaaggagaa gattcctgaa 840

atttgagaga caaaataaat ggaagacatg agaactttcc a 881

<210>55

<211>731

<212>DNA

<213> Intelligent people

<400>55

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcggacc cccccaagac acacgtgacc 120

caccaccctg tctttgacta tgaggccacc ctgaggtgct gggccctggg cttctaccct 180

gcggagatca tactgacctg gcagcgggat ggggaggacc agacccagga cgtggagctc 240

gtggagacca ggcctgcagg ggatggaacc ttccagaagt gggcagctgt ggtggtgcct 300

tctggagagg agcagagata cacgtgccat gtgcagcatg aggggctgcc ggagcccctc 360

atgctgagat ggatgtgaaa cagctgccct gtgtgggact gagtggcaag tccctttgtg 420

acttcaagaa ccctgactcc tctttgtgca gagaccagcc cacccctgtg cccaccatga 480

ccctcttcct catgctgaac tgcattcctt ccccaatcac ctttcctgtt ccagaaaagg 540

ggctgggatg tctccgtctc tgtctcaaat ttgtggtcca ctgagctata acttacttct 600

gtattaaaat tagaatctga gtataaattt actttttcaa attatttcca agagagattg 660

atgggttaat taaaggagaa gattcctgaa atttgagaga caaaataaat ggaagacatg 720

agaactttcc a 731

<210>56

<211>875

<212>DNA

<213> Intelligent people

<400>56

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggct cccactccat gaggtatttc 120

agcgccgccg tgtcccggcc cggccgcggg gagccccgct tcatcgccat gggctacgtg 180

gacgacacgc agttcgtgcg gttcgacagc gactcggcgt gtccgaggat ggagccgcgg 240

gcgccgtggg tggagcagga ggggccggag tattgggaag aggagacacg gaacaccaag 300

gcccacgcac agactgacag aatgaacctg cagaccctgc gcggctacta caaccagagc 360

gaggccaagc agtcttccct gcccaccatc cccatcatgg gtatcgttgc tggcctggtt 420

gtccttgcag ctgtagtcac tggagctgcg gtcgctgctg tgctgtggag aaagaagagc 480

tcagattgaa aaggagggag ctactctcag gctgcaatgt gaaacagctg ccctgtgtgg 540

gactgagtgg caagtccctt tgtgacttca agaaccctga ctcctctttg tgcagagacc 600

agcccacccc tgtgcccacc atgaccctct tcctcatgct gaactgcatt ccttccccaa 660

tcacctttcc tgttccagaa aaggggctgg gatgtctccg tctctgtctc aaatttgtgg 720

tccactgagc tataacttac ttctgtatta aaattagaat ctgagtataa atttactttt 780

tcaaattatt tccaagagag attgatgggt taattaaagg agaagattcc tgaaatttga 840

gagacaaaat aaatggaaga catgagaact ttcca 875

<210>57

<211>725

<212>DNA

<213> Intelligent people

<400>57

actcattctc cccagacgcc aaggatggtg gtcatggcgc cccgaaccct cttcctgctg 60

ctctcggggg ccctgaccct gaccgagacc tgggcgggct cccactccat gaggtatttc 120

agcgccgccg tgtcccggcc cggccgcggg gagccccgct tcatcgccat gggctacgtg 180

gacgacacgc agttcgtgcg gttcgacagc gactcggcgt gtccgaggat ggagccgcgg 240

gcgccgtggg tggagcagga ggggccggag tattgggaag aggagacacg gaacaccaag 300

gcccacgcac agactgacag aatgaacctg cagaccctgc gcggctacta caaccagagc 360

gaggccatgt gaaacagctg ccctgtgtgg gactgagtgg caagtccctt tgtgacttca 420

agaaccctga ctcctctttg tgcagagacc agcccacccc tgtgcccacc atgaccctct 480

tcctcatgct gaactgcatt ccttccccaa tcacctttcc tgttccagaa aaggggctgg 540

gatgtctccg tctctgtctc aaatttgtgg tccactgagc tataacttac ttctgtatta 600

aaattagaat ctgagtataa atttactttt tcaaattatt tccaagagag attgatgggt 660

taattaaagg agaagattcc tgaaatttga gagacaaaat aaatggaaga catgagaact 720

ttcca 725

<210>58

<211>1578

<212>DNA

<213> Intelligent people

<400>58

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggctcccact ccatgaggta tttcagcgcc gccgtgtccc ggcccggccg 300

cggggagccc cgcttcatcg ccatgggcta cgtggacgac acgcagttcg tgcggttcga 360

cagcgactcg gcgtgtccga ggatggagcc gcgggcgccg tgggtggagc aggaggggcc 420

ggagtattgg gaagaggaga cacggaacac caaggcccac gcacagactg acagaatgaa 480

cctgcagacc ctgcgcggct actacaacca gagcgaggcc agttctcaca ccctccagtg 540

gatgattggc tgcgacctgg ggtccgacgg acgcctcctc cgcgggtatg aacagtatgc 600

ctacgatggc aaggattacc tcgccctgaa cgaggacctg cgctcctgga ccgcagcgga 660

cactgcggct cagatctcca agcgcaagtg tgaggcggcc aatgtggctg aacaaaggag 720

agcctacctg gagggcacgt gcgtggagtg gctccacaga tacctggaga acgggaagga 780

gatgctgcag cgcgcggacc cccccaagac acacgtgacc caccaccctg tctttgacta 840

tgaggccacc ctgaggtgct gggccctggg cttctaccct gcggagatca tactgacctg 900

gcagcgggat ggggaggacc agacccagga cgtggagctc gtggagacca ggcctgcagg 960

ggatggaacc ttccagaagt gggcagctgt ggtggtgcct tctggagagg agcagagata 1020

cacgtgccat gtgcagcatg aggggctgcc ggagcccctc atgctgagat ggaagcagtc 1080

ttccctgccc accatcccca tcatgggtat cgttgctggc ctggttgtcc ttgcagctgt 1140

agtcactgga gctgcggtcg ctgctgtgct gtggagaaag aagagctcag attgaaaagg 1200

agggagctac tctcaggctg caatgtgaaa cagctgccct gtgtgggact gagtggcaag 1260

tccctttgtg acttcaagaa ccctgactcc tctttgtgca gagaccagcc cacccctgtg 1320

cccaccatga ccctcttcct catgctgaac tgcattcctt ccccaatcac ctttcctgtt 1380

ccagaaaagg ggctgggatg tctccgtctc tgtctcaaat ttgtggtcca ctgagctata 1440

acttacttct gtattaaaat tagaatctga gtataaattt actttttcaa attatttcca 1500

agagagattg atgggttaat taaaggagaa gattcctgaa atttgagaga caaaataaat 1560

ggaagacatg agaacttt 1578

<210>59

<211>1428

<212>DNA

<213> Intelligent people

<400>59

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggctcccact ccatgaggta tttcagcgcc gccgtgtccc ggcccggccg 300

cggggagccc cgcttcatcg ccatgggcta cgtggacgac acgcagttcg tgcggttcga 360

cagcgactcg gcgtgtccga ggatggagcc gcgggcgccg tgggtggagc aggaggggcc 420

ggagtattgg gaagaggaga cacggaacac caaggcccac gcacagactg acagaatgaa 480

cctgcagacc ctgcgcggct actacaacca gagcgaggcc agttctcaca ccctccagtg 540

gatgattggc tgcgacctgg ggtccgacgg acgcctcctc cgcgggtatg aacagtatgc 600

ctacgatggc aaggattacc tcgccctgaa cgaggacctg cgctcctgga ccgcagcgga 660

cactgcggct cagatctcca agcgcaagtg tgaggcggcc aatgtggctg aacaaaggag 720

agcctacctg gagggcacgt gcgtggagtg gctccacaga tacctggaga acgggaagga 780

gatgctgcag cgcgcggacc cccccaagac acacgtgacc caccaccctg tctttgacta 840

tgaggccacc ctgaggtgct gggccctggg cttctaccct gcggagatca tactgacctg 900

gcagcgggat ggggaggacc agacccagga cgtggagctc gtggagacca ggcctgcagg 960

ggatggaacc ttccagaagt gggcagctgt ggtggtgcct tctggagagg agcagagata 1020

cacgtgccat gtgcagcatg aggggctgcc ggagcccctc atgctgagat ggatgtgaaa 1080

cagctgccct gtgtgggact gagtggcaag tccctttgtg acttcaagaa ccctgactcc 1140

tctttgtgca gagaccagcc cacccctgtg cccaccatga ccctcttcct catgctgaac 1200

tgcattcctt ccccaatcac ctttcctgtt ccagaaaagg ggctgggatg tctccgtctc 1260

tgtctcaaat ttgtggtcca ctgagctata acttacttct gtattaaaat tagaatctga 1320

gtataaattt actttttcaa attatttcca agagagattg atgggttaat taaaggagaa 1380

gattcctgaa atttgagaga caaaataaat ggaagacatg agaacttt 1428

<210>60

<211>1308

<212>DNA

<213> Intelligent people

<400>60

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggttctcaca ccctccagtg gatgattggc tgcgacctgg ggtccgacgg 300

acgcctcctc cgcgggtatg aacagtatgc ctacgatggc aaggattacc tcgccctgaa 360

cgaggacctg cgctcctgga ccgcagcgga cactgcggct cagatctcca agcgcaagtg 420

tgaggcggcc aatgtggctg aacaaaggag agcctacctg gagggcacgt gcgtggagtg 480

gctccacaga tacctggaga acgggaagga gatgctgcag cgcgcggacc cccccaagac 540

acacgtgacc caccaccctg tctttgacta tgaggccacc ctgaggtgct gggccctggg 600

cttctaccct gcggagatca tactgacctg gcagcgggat ggggaggacc agacccagga 660

cgtggagctc gtggagacca ggcctgcagg ggatggaacc ttccagaagt gggcagctgt 720

ggtggtgcct tctggagagg agcagagata cacgtgccat gtgcagcatg aggggctgcc 780

ggagcccctc atgctgagat ggaagcagtc ttccctgccc accatcccca tcatgggtat 840

cgttgctggc ctggttgtcc ttgcagctgt agtcactgga gctgcggtcg ctgctgtgct 900

gtggagaaag aagagctcag attgaaaagg agggagctac tctcaggctg caatgtgaaa 960

cagctgccct gtgtgggact gagtggcaag tccctttgtg acttcaagaa ccctgactcc 1020

tctttgtgca gagaccagcc cacccctgtg cccaccatga ccctcttcct catgctgaac 1080

tgcattcctt ccccaatcac ctttcctgtt ccagaaaagg ggctgggatg tctccgtctc 1140

tgtctcaaat ttgtggtcca ctgagctata acttacttct gtattaaaat tagaatctga 1200

gtataaattt actttttcaa attatttcca agagagattg atgggttaat taaaggagaa 1260

gattcctgaa atttgagaga caaaataaat ggaagacatg agaacttt 1308

<210>61

<211>1158

<212>DNA

<213> Intelligent people

<400>61

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggttctcaca ccctccagtg gatgattggc tgcgacctgg ggtccgacgg 300

acgcctcctc cgcgggtatg aacagtatgc ctacgatggc aaggattacc tcgccctgaa 360

cgaggacctg cgctcctgga ccgcagcgga cactgcggct cagatctcca agcgcaagtg 420

tgaggcggcc aatgtggctg aacaaaggag agcctacctg gagggcacgt gcgtggagtg 480

gctccacaga tacctggaga acgggaagga gatgctgcag cgcgcggacc cccccaagac 540

acacgtgacc caccaccctg tctttgacta tgaggccacc ctgaggtgct gggccctggg 600

cttctaccct gcggagatcatactgacctg gcagcgggat ggggaggacc agacccagga 660

cgtggagctc gtggagacca ggcctgcagg ggatggaacc ttccagaagt gggcagctgt 720

ggtggtgcct tctggagagg agcagagata cacgtgccat gtgcagcatg aggggctgcc 780

ggagcccctc atgctgagat ggatgtgaaa cagctgccct gtgtgggact gagtggcaag 840

tccctttgtg acttcaagaa ccctgactcc tctttgtgca gagaccagcc cacccctgtg 900

cccaccatga ccctcttcct catgctgaac tgcattcctt ccccaatcac ctttcctgtt 960

ccagaaaagg ggctgggatg tctccgtctc tgtctcaaat ttgtggtcca ctgagctata 1020

acttacttct gtattaaaat tagaatctga gtataaattt actttttcaa attatttcca 1080

agagagattg atgggttaat taaaggagaa gattcctgaa atttgagaga caaaataaat 1140

ggaagacatg agaacttt 1158

<210>62

<211>1032

<212>DNA

<213> Intelligent people

<400>62

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggttctcaca ccctccagtg gatgattggc tgcgacctgg ggtccgacgg 300

acgcctcctc cgcgggtatg aacagtatgc ctacgatggc aaggattacc tcgccctgaa 360

cgaggacctg cgctcctgga ccgcagcgga cactgcggct cagatctcca agcgcaagtg 420

tgaggcggcc aatgtggctg aacaaaggag agcctacctg gagggcacgt gcgtggagtg 480

gctccacaga tacctggaga acgggaagga gatgctgcag cgcgcggagc agtcttccct 540

gcccaccatc cccatcatgg gtatcgttgc tggcctggtt gtccttgcag ctgtagtcac 600

tggagctgcg gtcgctgctg tgctgtggag aaagaagagc tcagattgaa aaggagggag 660

ctactctcag gctgcaatgt gaaacagctg ccctgtgtgg gactgagtgg caagtccctt 720

tgtgacttca agaaccctga ctcctctttg tgcagagacc agcccacccc tgtgcccacc 780

atgaccctct tcctcatgct gaactgcatt ccttccccaa tcacctttcc tgttccagaa 840

aaggggctgg gatgtctccg tctctgtctc aaatttgtgg tccactgagc tataacttac 900

ttctgtatta aaattagaat ctgagtataa atttactttt tcaaattatt tccaagagag 960

attgatgggt taattaaagg agaagattcc tgaaatttga gagacaaaat aaatggaaga 1020

catgagaact tt 1032

<210>63

<211>882

<212>DNA

<213> Intelligent people

<400>63

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggttctcaca ccctccagtg gatgattggc tgcgacctgg ggtccgacgg 300

acgcctcctc cgcgggtatg aacagtatgc ctacgatggc aaggattacc tcgccctgaa 360

cgaggacctg cgctcctgga ccgcagcgga cactgcggct cagatctcca agcgcaagtg 420

tgaggcggcc aatgtggctg aacaaaggag agcctacctg gagggcacgt gcgtggagtg 480

gctccacaga tacctggaga acgggaagga gatgctgcag cgcgcggtgt gaaacagctg 540

ccctgtgtgg gactgagtgg caagtccctt tgtgacttca agaaccctga ctcctctttg 600

tgcagagacc agcccacccc tgtgcccacc atgaccctct tcctcatgct gaactgcatt 660

ccttccccaa tcacctttcc tgttccagaa aaggggctgg gatgtctccg tctctgtctc 720

aaatttgtgg tccactgagc tataacttac ttctgtatta aaattagaat ctgagtataa 780

atttactttt tcaaattatt tccaagagag attgatgggt taattaaagg agaagattcc 840

tgaaatttga gagacaaaat aaatggaaga catgagaact tt 882

<210>64

<211>1032

<212>DNA

<213> Intelligent people

<400>64

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg gaccccccca agacacacgt gacccaccac cctgtctttg actatgaggc 300

caccctgagg tgctgggccc tgggcttcta ccctgcggag atcatactga cctggcagcg 360

ggatggggag gaccagaccc aggacgtgga gctcgtggag accaggcctg caggggatgg 420

aaccttccag aagtgggcag ctgtggtggt gccttctgga gaggagcaga gatacacgtg 480

ccatgtgcag catgaggggc tgccggagcc cctcatgctg agatggaagc agtcttccct 540

gcccaccatc cccatcatgg gtatcgttgc tggcctggtt gtccttgcag ctgtagtcac 600

tggagctgcg gtcgctgctg tgctgtggag aaagaagagc tcagattgaa aaggagggag 660

ctactctcag gctgcaatgt gaaacagctg ccctgtgtgg gactgagtgg caagtccctt 720

tgtgacttca agaaccctga ctcctctttg tgcagagacc agcccacccc tgtgcccacc 780

atgaccctct tcctcatgct gaactgcatt ccttccccaa tcacctttcc tgttccagaa 840

aaggggctgg gatgtctccg tctctgtctc aaatttgtgg tccactgagc tataacttac 900

ttctgtatta aaattagaat ctgagtataa atttactttt tcaaattatt tccaagagag 960

attgatgggt taattaaagg agaagattcc tgaaatttga gagacaaaat aaatggaaga 1020

catgagaact tt 1032

<210>65

<211>882

<212>DNA

<213> Intelligent people

<400>65

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg gaccccccca agacacacgt gacccaccac cctgtctttg actatgaggc 300

caccctgagg tgctgggccc tgggcttcta ccctgcggag atcatactga cctggcagcg 360

ggatggggag gaccagaccc aggacgtgga gctcgtggag accaggcctg caggggatgg 420

aaccttccag aagtgggcag ctgtggtggt gccttctgga gaggagcaga gatacacgtg 480

ccatgtgcag catgaggggc tgccggagcc cctcatgctg agatggatgt gaaacagctg 540

ccctgtgtgg gactgagtgg caagtccctt tgtgacttca agaaccctga ctcctctttg 600

tgcagagacc agcccacccc tgtgcccacc atgaccctct tcctcatgct gaactgcatt 660

ccttccccaa tcacctttcc tgttccagaa aaggggctgg gatgtctccg tctctgtctc 720

aaatttgtgg tccactgagc tataacttac ttctgtatta aaattagaat ctgagtataa 780

atttactttt tcaaattatt tccaagagag attgatgggt taattaaagg agaagattcc 840

tgaaatttga gagacaaaat aaatggaaga catgagaact tt 882

<210>66

<211>1026

<212>DNA

<213> Intelligent people

<400>66

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggctcccact ccatgaggta tttcagcgcc gccgtgtccc ggcccggccg 300

cggggagccc cgcttcatcg ccatgggcta cgtggacgac acgcagttcg tgcggttcga 360

cagcgactcg gcgtgtccga ggatggagcc gcgggcgccg tgggtggagc aggaggggcc 420

ggagtattgg gaagaggaga cacggaacac caaggcccac gcacagactg acagaatgaa 480

cctgcagacc ctgcgcggct actacaacca gagcgaggcc aagcagtctt ccctgcccac 540

catccccatc atgggtatcg ttgctggcct ggttgtcctt gcagctgtag tcactggagc 600

tgcggtcgct gctgtgctgt ggagaaagaa gagctcagat tgaaaaggag ggagctactc 660

tcaggctgca atgtgaaaca gctgccctgt gtgggactga gtggcaagtc cctttgtgac 720

ttcaagaacc ctgactcctc tttgtgcaga gaccagccca cccctgtgcc caccatgacc 780

ctcttcctca tgctgaactg cattccttcc ccaatcacct ttcctgttcc agaaaagggg 840

ctgggatgtc tccgtctctg tctcaaattt gtggtccact gagctataac ttacttctgt 900

attaaaatta gaatctgagt ataaatttac tttttcaaat tatttccaag agagattgat 960

gggttaatta aaggagaaga ttcctgaaat ttgagagaca aaataaatgg aagacatgag 1020

aacttt 1026

<210>67

<211>876

<212>DNA

<213> Intelligent people

<400>67

agtgtggtac tttgtcttga ggagatgtcc tggactcaca cggaaactta gggctacgga 60

atgaagttct cactcccatt aggtgacagg tttttagaga agccaatcag cgtcgccgcg 120

gtcctggttc taaagtcctc gctcacccac ccggactcat tctccccaga cgccaaggat 180

ggtggtcatg gcgccccgaa ccctcttcct gctgctctcg ggggccctga ccctgaccga 240

gacctgggcg ggctcccact ccatgaggta tttcagcgcc gccgtgtccc ggcccggccg 300

cggggagccc cgcttcatcg ccatgggcta cgtggacgac acgcagttcg tgcggttcga 360

cagcgactcg gcgtgtccga ggatggagcc gcgggcgccg tgggtggagc aggaggggcc 420

ggagtattgg gaagaggaga cacggaacac caaggcccac gcacagactg acagaatgaa 480

cctgcagacc ctgcgcggct actacaacca gagcgaggcc atgtgaaaca gctgccctgt 540

gtgggactga gtggcaagtc cctttgtgac ttcaagaacc ctgactcctc tttgtgcaga 600

gaccagccca cccctgtgcc caccatgacc ctcttcctca tgctgaactg cattccttcc 660

ccaatcacct ttcctgttcc agaaaagggg ctgggatgtc tccgtctctg tctcaaattt 720

gtggtccact gagctataac ttacttctgt attaaaatta gaatctgagt ataaatttac 780

tttttcaaat tatttccaag agagattgat gggttaatta aaggagaaga ttcctgaaat 840

ttgagagaca aaataaatgg aagacatgag aacttt 876

<210>68

<211>781

<212>DNA

<213> Intelligent people

<400>68

gttctcacac cctccagtgg atgattggct gcgacctggg gtccgacgga cgcctcctcc 60

gcgggtatga acagtatgcc tacgatggca aggattacct cgccctgaac gaggacctgc 120

gctcctggac cgcagcggac actgcggctc agatctccaa gcgcaagtgt gaggcggcca 180

atgtggctga acaaaggaga gcctacctgg agggcacgtg cgtggagtgg ctccacagat 240

acctggagaa cgggaaggag atgctgcagc gcgcggagca gtcttccctg cccaccatcc 300

ccatcatggg tatcgttgct ggcctggttg tccttgcagc tgtagtcact ggagctgcgg 360

tcgctgctgt gctgtggaga aagaagagct cagattgaaa aggagggagc tactctcagg 420

ctgcaatgtg aaacagctgc cctgtgtggg actgagtggc aagtcccttt gtgacttcaa 480

gaaccctgac tcctctttgt gcagagacca gcccacccct gtgcccacca tgaccctctt 540

cctcatgctg aactgcattc cttccccaat cacctttcct gttccagaaa aggggctggg 600

atgtctccgt ctctgtctca aatttgtggt ccactgagct ataacttact tctgtattaa 660

aattagaatc tgagtataaa tttacttttt caaattattt ccaagagaga ttgatgggtt 720

aattaaagga gaagattcct gaaatttgag agacaaaata aatggaagac atgagaactt 780

t 781

<210>69

<211>631

<212>DNA

<213> Intelligent people

<400>69

gttctcacac cctccagtgg atgattggct gcgacctggg gtccgacgga cgcctcctcc 60

gcgggtatga acagtatgcc tacgatggca aggattacct cgccctgaac gaggacctgc 120

gctcctggac cgcagcggac actgcggctc agatctccaa gcgcaagtgt gaggcggcca 180

atgtggctga acaaaggaga gcctacctgg agggcacgtg cgtggagtgg ctccacagat 240

acctggagaa cgggaaggag atgctgcagc gcgcggtgtg aaacagctgc cctgtgtggg 300

actgagtggc aagtcccttt gtgacttcaa gaaccctgac tcctctttgt gcagagacca 360

gcccacccct gtgcccacca tgaccctctt cctcatgctg aactgcattc cttccccaat 420

cacctttcct gttccagaaa aggggctggg atgtctccgt ctctgtctca aatttgtggt 480

ccactgagct ataacttact tctgtattaa aattagaatc tgagtataaa tttacttttt 540

caaattattt ccaagagaga ttgatgggtt aattaaagga gaagattcct gaaatttgag 600

agacaaaata aatggaagac atgagaactt t 631

<210>70

<211>1057

<212>DNA

<213> Intelligent people

<400>70

gttctcacac cctccagtgg atgattggct gcgacctggg gtccgacgga cgcctcctcc 60

gcgggtatga acagtatgcc tacgatggca aggattacct cgccctgaac gaggacctgc 120

gctcctggac cgcagcggac actgcggctc agatctccaa gcgcaagtgt gaggcggcca 180

atgtggctga acaaaggaga gcctacctgg agggcacgtg cgtggagtgg ctccacagat 240

acctggagaa cgggaaggag atgctgcagc gcgcggaccc ccccaagaca cacgtgaccc 300

accaccctgt ctttgactat gaggccaccc tgaggtgctg ggccctgggc ttctaccctg 360

cggagatcat actgacctgg cagcgggatg gggaggacca gacccaggac gtggagctcg 420

tggagaccag gcctgcaggg gatggaacct tccagaagtg ggcagctgtg gtggtgcctt 480

ctggagagga gcagagatac acgtgccatg tgcagcatga ggggctgccg gagcccctca 540

tgctgagatg gaagcagtct tccctgccca ccatccccat catgggtatc gttgctggcc 600

tggttgtcct tgcagctgta gtcactggag ctgcggtcgc tgctgtgctg tggagaaaga 660

agagctcaga ttgaaaagga gggagctact ctcaggctgc aatgtgaaac agctgccctg 720

tgtgggactg agtggcaagt ccctttgtga cttcaagaac cctgactcct ctttgtgcag 780

agaccagccc acccctgtgc ccaccatgac cctcttcctc atgctgaact gcattccttc 840

cccaatcacc tttcctgttc cagaaaaggg gctgggatgt ctccgtctct gtctcaaatt 900

tgtggtccac tgagctataa cttacttctg tattaaaatt agaatctgag tataaattta 960

ctttttcaaa ttatttccaa gagagattga tgggttaatt aaaggagaag attcctgaaa 1020

tttgagagac aaaataaatg gaagacatga gaacttt 1057

<210>71

<211>907

<212>DNA

<213> Intelligent people

<400>71

gttctcacac cctccagtgg atgattggct gcgacctggg gtccgacgga cgcctcctcc 60

gcgggtatga acagtatgcc tacgatggca aggattacct cgccctgaac gaggacctgc 120

gctcctggac cgcagcggac actgcggctc agatctccaa gcgcaagtgt gaggcggcca 180

atgtggctga acaaaggaga gcctacctgg agggcacgtg cgtggagtgg ctccacagat 240

acctggagaa cgggaaggag atgctgcagc gcgcggaccc ccccaagaca cacgtgaccc 300

accaccctgt ctttgactat gaggccaccc tgaggtgctg ggccctgggc ttctaccctg 360

cggagatcat actgacctgg cagcgggatg gggaggacca gacccaggac gtggagctcg 420

tggagaccag gcctgcaggg gatggaacct tccagaagtg ggcagctgtg gtggtgcctt 480

ctggagagga gcagagatac acgtgccatg tgcagcatga ggggctgccg gagcccctca 540

tgctgagatg gatgtgaaac agctgccctg tgtgggactg agtggcaagt ccctttgtga 600

cttcaagaac cctgactcct ctttgtgcag agaccagccc acccctgtgc ccaccatgac 660

cctcttcctc atgctgaact gcattccttc cccaatcacc tttcctgttc cagaaaaggg 720

gctgggatgt ctccgtctct gtctcaaatt tgtggtccac tgagctataa cttacttctg 780

tattaaaatt agaatctgag tataaattta ctttttcaaa ttatttccaa gagagattga 840

tgggttaatt aaaggagaag attcctgaaa tttgagagac aaaataaatg gaagacatga 900

gaacttt 907

<210>72

<211>806

<212>DNA

<213> Intelligent people

<400>72

gggaaggaga tgctgcagcg cgcggacccc cccaagacac acgtgaccca ccaccctgtc 60

tttgactatg aggccaccct gaggtgctgg gccctgggct tctaccctgc ggagatcata 120

ctgacctggc agcgggatgg ggaggaccag acccaggacg tggagctcgt ggagaccagg 180

cctgcagggg atggaacctt ccagaagtgg gcagctgtgg tggtgccttc tggagaggag 240

cagagataca cgtgccatgt gcagcatgag gggctgccgg agcccctcat gctgagatgg 300

aagcagtctt ccctgcccac catccccatc atgggtatcg ttgctggcct ggttgtcctt 360

gcagctgtag tcactggagc tgcggtcgct gctgtgctgt ggagaaagaa gagctcagat 420

tgaaaaggag ggagctactc tcaggctgca atgtgaaaca gctgccctgt gtgggactga 480

gtggcaagtc cctttgtgac ttcaagaacc ctgactcctc tttgtgcaga gaccagccca 540

cccctgtgcc caccatgacc ctcttcctca tgctgaactg cattccttcc ccaatcacct 600

ttcctgttcc agaaaagggg ctgggatgtc tccgtctctg tctcaaattt gtggtccact 660

gagctataac ttacttctgt attaaaatta gaatctgagt ataaatttac tttttcaaat 720

tatttccaag agagattgat gggttaatta aaggagaaga ttcctgaaat ttgagagaca 780

aaataaatgg aagacatgag aacttt 806

<210>73

<211>781

<212>DNA

<213> Intelligent people

<400>73

acccccccaa gacacacgtg acccaccacc ctgtctttga ctatgaggcc accctgaggt 60

gctgggccct gggcttctac cctgcggaga tcatactgac ctggcagcgg gatggggagg 120

accagaccca ggacgtggag ctcgtggaga ccaggcctgc aggggatgga accttccaga 180

agtgggcagc tgtggtggtg ccttctggag aggagcagag atacacgtgc catgtgcagc 240

atgaggggct gccggagccc ctcatgctga gatggaagca gtcttccctg cccaccatcc 300

ccatcatggg tatcgttgct ggcctggttg tccttgcagc tgtagtcact ggagctgcgg 360

tcgctgctgt gctgtggaga aagaagagct cagattgaaaaggagggagc tactctcagg 420

ctgcaatgtg aaacagctgc cctgtgtggg actgagtggc aagtcccttt gtgacttcaa 480

gaaccctgac tcctctttgt gcagagacca gcccacccct gtgcccacca tgaccctctt 540

cctcatgctg aactgcattc cttccccaat cacctttcct gttccagaaa aggggctggg 600

atgtctccgt ctctgtctca aatttgtggt ccactgagct ataacttact tctgtattaa 660

aattagaatc tgagtataaa tttacttttt caaattattt ccaagagaga ttgatgggtt 720

aattaaagga gaagattcct gaaatttgag agacaaaata aatggaagac atgagaactt 780

t 781

<210>74

<211>656

<212>DNA

<213> Intelligent people

<400>74

gggaaggaga tgctgcagcg cgcggacccc cccaagacac acgtgaccca ccaccctgtc 60

tttgactatg aggccaccct gaggtgctgg gccctgggct tctaccctgc ggagatcata 120

ctgacctggc agcgggatgg ggaggaccag acccaggacg tggagctcgt ggagaccagg 180

cctgcagggg atggaacctt ccagaagtgg gcagctgtgg tggtgccttc tggagaggag 240

cagagataca cgtgccatgt gcagcatgag gggctgccgg agcccctcat gctgagatgg 300

atgtgaaaca gctgccctgt gtgggactga gtggcaagtc cctttgtgac ttcaagaacc 360

ctgactcctc tttgtgcaga gaccagccca cccctgtgcc caccatgacc ctcttcctca 420

tgctgaactg cattccttcc ccaatcacct ttcctgttcc agaaaagggg ctgggatgtc 480

tccgtctctg tctcaaattt gtggtccact gagctataac ttacttctgt attaaaatta 540

gaatctgagt ataaatttac tttttcaaat tatttccaag agagattgat gggttaatta 600

aaggagaaga ttcctgaaat ttgagagaca aaataaatgg aagacatgag aacttt 656

<210>75

<211>16

<212>DNA

<213> Artificial

<220>

<223> primer

<400>75

ccactgcccc tggtac 16

<210>76

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>76

cctggactca cacggaaact 20

<210>77

<211>21

<212>DNA

<213> Artificial

<220>

<223> primer

<400>77

gtaacatagt gtggtacttt g 21

<210>78

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>78

cccaaggcgc ctttaccaaa 20

<210>79

<211>17

<212>DNA

<213> Artificial

<220>

<223> primer

<400>79

agccctcacc accgacc 17

<210>80

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>80

cctttgttca gccacattgg 20

<210>81

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>81

ggactcattc tccccagacg 20

<210>82

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>82

ggaagaggag acacggaaca 20

<210>83

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>83

tgttccgtgt ctcctcttcc 20

<210>84

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>84

ccaatgtggc tgaacaaagg 20

<210>85

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>85

cctttgttca gccacattgg 20

<210>86

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>86

gcagctccag tgactacagc 20

<210>87

<211>21

<212>DNA

<213> Artificial

<220>

<223> primer

<400>87

tcctgtggca tccacgaaac t 21

<210>88

<211>21

<212>DNA

<213> Artificial

<220>

<223> primer

<400>88

gaagcatttg cggtggacga t 21

<210>89

<211>20

<212>DNA

<213> Artificial

<220>

<223> primer

<400>89

ggcctcaagc gtggctctca 20

<210>90

<211>319

<212>PRT

<213> Intelligent people

<400>90

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Ser Met Arg Tyr Phe

20 25 30

Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu Pro Arg Phe Ile Ala

35 40 45

Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ser

50 55 60

Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp Val Glu Gln Glu Gly

65 70 75 80

Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr Lys Ala His Ala Gln

85 90 95

Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly Tyr Tyr Asn Gln Ser

100 105 110

Glu Ala Ser Ser His Thr Leu Gln Trp Met Ile Gly Cys Asp Leu Gly

115 120 125

Ser Asp Gly Arg Leu Leu Arg Gly Tyr Glu Gln Tyr Ala Tyr Asp Gly

130 135 140

Lys AspTyr Leu Ala Leu Asn Glu Asp Leu Arg Ser Trp Thr Ala Ala

145 150 155 160

Asp Thr Ala Ala Gln Ile Ser Lys Arg Lys Cys Glu Ala Ala Asn Val

165 170 175

Ala Glu Gln Arg Arg Ala Tyr Leu Glu Gly Thr Cys Val Glu Trp Leu

180 185 190

His Arg Tyr Leu Glu Asn Gly Lys Glu Met Leu Gln Arg Ala Asp Pro

195 200 205

Pro Lys Thr His Val Thr His His Pro Val Phe Asp Tyr Glu Ala Thr

210 215 220

Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu Ile Ile Leu Thr

225 230 235 240

Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val Glu Leu Val Glu

245 250 255

Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp Ala Ala Val Val

260 265 270

Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His Val Gln His Glu

275 280 285

Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Ser Lys Glu Gly Asp Gly

290 295 300

Gly Ile Met SerVal Arg Glu Ser Arg Ser Leu Ser Glu Asp Leu

305 310 315

<210>91

<211>227

<212>PRT

<213> Intelligent people

<400>91

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Ser Met Arg Tyr Phe

20 25 30

Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu Pro Arg Phe Ile Ala

35 40 45

Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ser

50 55 60

Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp Val Glu Gln Glu Gly

65 70 75 80

Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr Lys Ala His Ala Gln

85 90 95

Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly Tyr Tyr Asn Gln Ser

100 105 110

Glu Ala Asn Pro Pro Lys Thr His Val Thr His His Pro Val Phe Asp

115 120 125

Tyr Glu Ala Thr Leu Arg Cys Trp Ala Leu Gly Phe Tyr Pro Ala Glu

130 135 140

Ile Ile Leu Thr Trp Gln Arg Asp Gly Glu Asp Gln Thr Gln Asp Val

145 150 155 160

Glu Leu Val Glu Thr Arg Pro Ala Gly Asp Gly Thr Phe Gln Lys Trp

165 170 175

Ala Ala Val Val Val Pro Ser Gly Glu Glu Gln Arg Tyr Thr Cys His

180 185 190

Val Gln His Glu Gly Leu Pro Glu Pro Leu Met Leu Arg Trp Ser Lys

195 200 205

Glu Gly Asp Gly Gly Ile Met Ser Val Arg Glu Ser Arg Ser Leu Ser

210 215 220

Glu Asp Leu

225

<210>92

<211>116

<212>PRT

<213> Intelligent people

<400>92

Met Val Val Met Ala Pro Arg Thr Leu Phe Leu Leu Leu Ser Gly Ala

1 5 10 15

Leu Thr Leu Thr Glu Thr Trp Ala Gly Ser His Ser Met Arg Tyr Phe

20 25 30

Ser Ala Ala Val Ser Arg Pro Gly Arg Gly Glu Pro Arg Phe Ile Ala

35 40 45

Met Gly Tyr Val Asp Asp Thr Gln Phe Val Arg Phe Asp Ser Asp Ser

50 55 60

Ala Cys Pro Arg Met Glu Pro Arg Ala Pro Trp Val Glu Gln Glu Gly

65 70 75 80

Pro Glu Tyr Trp Glu Glu Glu Thr Arg Asn Thr Lys Ala His Ala Gln

85 90 95

Thr Asp Arg Met Asn Leu Gln Thr Leu Arg Gly Tyr Tyr Asn Gln Ser

100 105 110

Glu Ala Ser Glu

115

<210>93

<211>13

<212>DNA

<213> Intelligent people

<400>93

agtgtggtac ttt 13

<210>94

<211>22

<212>DNA

<213> Intelligent people

<400>94

tggaagacat gagaactttc ca 22

<210>95

<211>17

<212>DNA

<213> Intelligent people

<400>95

atatagtaac atagtgt 17