C-糖基转移酶及其在合成夏佛塔苷和异夏佛塔苷中的应用
阅读说明：本技术 C-糖基转移酶及其在合成夏佛塔苷和异夏佛塔苷中的应用 (C-glycosyltransferase and application thereof in synthesis of schaftoside and isoschaftoside ) 是由 乔雪 叶敏 王子龙 王双 陈宽 于 2019-04-10 设计创作，主要内容包括：本发明公开了一种C-糖基转移酶及其在合成夏佛塔苷和异夏佛塔苷中的应用。本发明从黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中分离获得了2-羟基黄烷酮C-糖基转移酶基因(SbCGTa、GuCGTa、PsCGTa、LpCGTa、AeCGTa)和C-葡萄糖-2-羟基黄烷酮C-糖基转移酶基因(SbCGTb、ZmCGTb、OsCGTb1、OsCGTb2、GuCGTb、PsCGTb、LpCGTb、AeCGTb),首次发现这些基因编码的蛋白参与合成植物中植保素夏佛塔苷和异夏佛塔苷,从而为体外利用酶催化方法合成夏佛塔苷、异夏佛塔苷及其类似物提供了可行的方法。(The invention discloses C-glycosyltransferase and application thereof in synthesizing schaftoside and isoschaftoside. The invention separates 2-hydroxyflavanone C-glycosyltransferase genes (SbCGTa, GuCGTa, PsCGTa, LpCGTa, AeCGTa) and C-glucose-2-hydroxyflavanone C-glycosyltransferase genes (SbCGTb, ZmCGTb, OsCGTb1, OsCGTb2, GuCGTb, PsCGTb, LpCGTb and AeCGTb) from scutellaria baicalensis, corn, rice, liquorice, pistia stratiotes, duckweed and arisaema sylvestre, and discovers that proteins coded by the genes participate in the synthesis of phytoalexin schaftoside and isoschaftoside in plants for the first time, thereby providing a feasible method for synthesizing schaftoside, isoschaftoside and analogues thereof by an enzyme catalysis method in vitro.)
技术领域
本发明涉及分子生物学领域，尤其涉及C-糖基转移酶(CGT)基因及其编码蛋白和在合成夏佛塔苷和异夏佛塔苷中的应用。
背景技术
夏佛塔苷和异夏佛塔苷是一类具有不同糖基的黄酮双碳苷，作为植保素，在多种植物中均有分布。目前在19个科39种植物中含有该类成分，包括水生和陆生植物、单子叶和双子叶植物、以及草木和灌木植物中1。夏佛塔苷和异夏佛塔苷具有明显的生理学活性和药理学活性。在金线草中，夏佛塔苷作为异感化物，可抑制独脚金生长2a-c。在水稻的韧皮部中，夏佛塔苷作为拒食剂。青蒿块茎中的夏佛塔苷和异夏佛塔苷具有杀线虫活性。此外，夏佛塔苷和异夏佛塔苷也具有多种药理活性。芋头、硬核属植物(Scleropyrum pentandrum)、甘蔗糖蜜和黄芩中的夏佛塔苷和异夏佛塔苷表现出抗氧化活性3a-d。金钱草中的夏佛塔苷作为血管紧张素转化酶抑制剂表现出抗高血压功能4。合果芋中的异夏佛塔苷通过降低肾Na+,K+-ATP酶活性表现出抗高血压活性5。闭鞘姜属植物(Costus spiralis)叶中的夏佛塔苷和异夏佛塔苷通过抑制α-糖苷酶表现出抗糖尿病功能6。麻风树叶中的异夏佛塔苷对A375细胞表现出抗恶性细胞增生的活性7。异叶杜鹃和牛筋草中的夏佛塔苷具有抗肝毒性和抑制LPS诱导的小鼠肺炎活性8a-b。然而，夏佛塔苷和异夏佛塔苷在植物中的生物合成过程却不清楚。
近年来，关于植物中黄酮碳苷的生物合成途径也有报道，主要有三种：1)水稻中C-糖基转移酶(OsCGT)催化2-羟基黄烷酮生成C-葡萄糖-2-羟基黄烷酮，经脱水酶选择性催化生成黄酮-6-C-葡萄糖，如异牡荆苷和异荭草素9。此外，玉米中C-糖基转移酶(UGT708A6)10、荞麦中C-糖基转移酶(FeCGTa和FeCGTb)11、大豆中C-糖基转移酶(UGT708D1)12的催化功能与OsCGT的催化功能相似。以上C-糖基转移酶除了催化功能相似，在氨基酸序列上也有一定的相似性，均含有DPFF保守区。2)龙胆中C-糖基转移酶(GtUF6CGT1)直接催化芹菜素生成异牡荆苷，这是一种直接在黄酮母核上催化生成黄酮单葡萄糖碳苷的反应13。GtUF6CGT1选择性在黄酮C-6位进行碳苷化反应。相似的，葛根中C-糖基转移酶(PlUGT43)直接催化大豆素生成葛根素14。PlUGT43选择性在异黄酮的C-8位进行碳苷化反应。3)对于黄酮双葡萄糖碳苷的生物合成，仅发现橘属中2个C-糖基转移酶(FcCGT和CuCGT)15。FcCGT和CuCGT先催化2-羟基黄烷酮生成C-葡萄糖-2-羟基黄烷酮，再进一步催化生成2C-葡萄糖-2-羟基黄烷酮，脱水产生新西兰牡荆苷。FcCGT和CuCGT与大部分植物中的C-糖基转移酶相似，也具有DPFF保守区。然而，具有不同糖基的黄酮双碳苷，如夏佛塔苷和异夏佛塔苷的生物合成过程尚不清楚。因此，阐明植物中植保素夏佛塔苷和异夏佛塔苷的生物合成过程具有重要的意义。
发明内容
本发明的目的在于阐明夏佛塔苷和异夏佛塔苷的生物合成过程，并建立了体外酶法合成夏佛塔苷、异夏佛塔苷及其类似物的方法。
为实现上述技术目的，本发明通过转录组分析的方法，从黄芩(Scutellariabaicalensis)、玉米(Zea mays)、水稻(Oryza sativa)、甘草(Glycyrrhiza uralensis)、大薸(Pistia stratiotes)、少根紫萍(Landoltia punctate)、一把伞南星(Arisaemaerubescens)七种植物中筛选出可能的C-糖基转移酶基因序列。
首先，采用RT-PCR的方法扩增可能的C-糖基转移酶基因，经改良的quick-change法或无缝拼接的方法构建重组载体，并在大肠杆菌中表达，经纯化后获得相应的C-糖基转移酶蛋白。
通过功能表征，在黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中共发现13个C-糖基转移酶，即五个2-羟基黄烷酮C-糖基转移酶(SbCGTa、GuCGTa、PsCGTa、LpCGTa、AeCGTa)和八个C-葡萄糖-2-羟基黄烷酮C-糖基转移酶(SbCGTb、ZmCGTb、OsCGTb1、OsCGTb2、GuCGTb、PsCGTb、LpCGTb、AeCGTb)。
从黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中分离获得的五种2-羟基黄烷酮C-糖基转移酶基因分别命名为SbCGTa、GuCGTa、PsCGTa、LpCGTa、AeCGTa，它们的核苷酸序列分别如序列表中SEQ ID No：1、5、11、9和7所示；八种C-葡萄糖-2-羟基黄烷酮C-糖基转移酶基因分别命名为SbCGTb、ZmCGTb、OsCGTb1、OsCGTb2、GuCGTb、PsCGTb、LpCGTb和AeCGTb，其核苷酸序列分别如序列表中SEQ ID No：2、13、3、4、6、12、10和8所示。
含有上述2-羟基黄烷酮C-糖基转移酶基因或C-葡萄糖-2-羟基黄烷酮C-糖基转移酶基因的载体、表达盒、宿主菌和转基因细胞系也在本发明的保护范围内。
上述2-羟基黄烷酮C-糖基转移酶基因编码的蛋白SbCGTa、GuCGTa、AeCGTa、LpCGTa、和PsCGTa的氨基酸序列分别如序列表中SEQ ID No：14、18、20、22和24所示。上述C-葡萄糖-2-羟基黄烷酮C-糖基转移酶基因编码的蛋白ZmCGTb、OsCGTb1、OsCGTb2、SbCGTb、GuCGTb、AeCGTb、LpCGTb、PsCGTb的氨基酸序列分别如序列表中SEQ ID No：26、16、17、15、19、21、23和25所示。
本发明发现，上述C-糖基转移酶参与植物中植保素夏佛塔苷和异夏佛塔苷的生物合成过程。利用上述2-羟基黄烷酮C-糖基转移酶和C-葡萄糖-2-羟基黄烷酮C-糖基转移酶通过体外酶法可以实现夏佛塔苷、异夏佛塔苷、白杨素-6-C-α-L-***糖-8-C-β-D-葡萄糖、白杨素-6-C-β-D-葡萄糖-8-C-α-L-***糖、3’-C-葡萄糖-5’-C-***糖根皮素等类似物的合成。
由此，本发明提供了一种合成葡萄糖和***糖双糖基碳苷化合物的方法，包括：利用2-羟基黄烷酮C-糖基转移酶催化式I化合物和尿苷二磷酸葡萄糖反应生成式II化合物，再利用C-葡萄糖-2-羟基黄烷酮C-糖基转移酶催化式II化合物和尿苷二磷酸***糖反应生成式III化合物，如下所示：
其中，R1和R2可以相互独立或者R1和R2成环；
当R1和R2相互独立时，R1为H、羟基或烷氧基，R2为H、烷基、烷氧基、苯基、取代苯基、苯基亚烷基、取代苯基亚烷基、苯氧基、取代苯氧基、苯基烷氧基、取代苯基烷氧基、苯乙酮基、对羟基苯乙酮基、3’,4’-二羟基苯乙酮基；
或者，在式I中R1和R2成环，所述式I化合物结构如下：
上式中，R为H或OH；相应得到的式II化合物结构如下：
上式中，R为H或OH；相应得到的式III化合物结构如下：
当R1为烷氧基时，所述烷氧基优选为C1～C6烷氧基，例如甲氧基(-OCH3)、乙氧基(-OCH2CH3)和丙氧基(-OCH2CH2CH3)等。
当R2为烷基时，所述烷基优选为C1-C10的烷基，更优选为C1-C6的烷基，例如甲基、乙基、丙基、异丙基等。
当R2为烷氧基时，所述烷氧基优选为C1～C6烷氧基，例如甲氧基(-OCH3)、乙氧基(-OCH2CH3)和丙氧基(-OCH2CH2CH3)等。
当R2为取代苯基时，苯基上的取代基可以是一个或多个，这些取代基可以在对位、间位或邻位，所述取代基优选为羟基、C1-C6的烷基、C1-C6的烷氧基，例如甲基、乙基、丙基、甲氧基、乙氧基、丙氧基等。
当R2为苯基亚烷基时，可表示为-(CH2)n-ph，例如为苯基亚甲基(即苄基)、苯基亚乙基等；当R2为取代苯基亚烷基时，可表示为-(CH2)n-取代苯基，所述取代苯基上的取代基可以是一个或多个，这些取代基可以在对位、间位或邻位，所述取代基优选为羟基、C1-C6的烷基、C1-C6的烷氧基，例如甲基、乙基、丙基、甲氧基、乙氧基、丙氧基等。其中n为正整数，优选为1～6的正整数。
当R2为苯氧基、取代苯氧基、苯基烷氧基、取代苯基烷氧基、苯乙酮基、对羟基苯乙酮基、3’,4’-二羟基苯乙酮基时，其结构如下所示：
上述取代苯氧基中苯基上的R3代表一个或多个取代基，这些取代基可以在对位、间位或邻位，所述取代基优选为羟基、C1-C6的烷基、C1-C6的烷氧基，例如甲基、乙基、丙基、甲氧基、乙氧基、丙氧基等。
上述取代苯基烷氧基中苯基上的R4代表一个或多个取代基，这些取代基可以在对位、间位或邻位，所述取代基优选为羟基、C1-C6的烷基、C1-C6的烷氧基，例如甲基、乙基、丙基、甲氧基、乙氧基、丙氧基等。
上述苯基烷氧基和取代苯基烷氧基结构式中的n代表正整数，优选为1～6的正整数。
进一步优选的，R1为氢、羟基或甲氧基；R2选自下列基团之一：
参见图1，合成夏佛塔苷、异夏佛塔苷或其类似物的体外酶催化反应如下：2-羟基黄烷酮C-糖基转移酶(CGTa)以2-羟基黄烷酮和尿苷二磷酸葡萄糖(UDPG)为原料，催化生成C-葡萄糖-2-羟基黄烷酮；C-葡萄糖-2-羟基黄烷酮C-糖基转移酶(CGTb)以C-葡萄糖-2-羟基黄烷酮和尿苷二磷酸***糖(UDP-ara)为原料，催化生成C-***糖-C-葡萄糖-2-羟基黄烷酮；C-***糖-C-葡萄糖-2-羟基黄烷酮经HCl处理脱水或自发脱水生成夏佛塔苷、异夏佛塔苷、白杨素-6-C-α-L-***糖-8-C-β-D-葡萄糖、白杨素-6-C-β-D-葡萄糖-8-C-α-L-***糖等类似物同分异构体。
本发明的有益效果：
本发明首次发现了黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中13个C-糖基转移酶。这些酶参与合成植物中植保素夏佛塔苷和异夏佛塔苷，阐明其生物合成过程。本发明公布了体外利用酶催化方法，为合成夏佛塔苷、异夏佛塔苷及其类似物，提供了可行的方法。
附图说明
图1.利用本发明的2-羟基黄烷酮C-糖基转移酶和C-葡萄糖-2-羟基黄烷酮C-糖基转移酶体外酶法合成夏佛塔苷和异夏佛塔苷及其类似物的反应式。
图2.SbCGTa和SbCGTb依次催化2-羟基松属素(1)生成白杨素-6-C-α-L-***糖-8-C-β-D-葡萄糖(5a)和白杨素-6-C-β-D-葡萄糖-8-C-α-L-***糖(5b)，SbCGTa和SbCGTb依次催化2-羟基柚皮素(2)生成夏佛塔苷(6b)、异夏佛塔苷(6a)的反应式和各化合物的液相色谱图。
图3.SbCGTa和SbCGTb催化根皮素(7)生成3’-C-葡萄糖-5’-C-***糖根皮素(9)的反应式和各化合物的液相色谱图。
图4.SbCGTa和SbCGTb依次催化夫洛丙酮(10)生成3’-C-葡萄糖-5’-C-***糖夫洛丙酮(14)，SbCGTa和SbCGTb依次催化2,4,6-三羟基苯甲酸甲酯(11)生成2,4,6-三羟基-3-C-葡萄糖-5-C-***糖苯甲酸甲酯(15)的反应式和各化合物的液相色谱图。
图5.玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中2-羟基黄烷酮C-糖基转移酶(CGTa)和C-葡萄糖-2-羟基黄烷酮C-糖基转移酶(CGTb)的功能表征实验结果，其中：
(A)CGTa催化2-羟基柚皮素生成C-葡萄糖-2-羟基柚皮素，CGTa催化2-羟基松属素生成C-葡萄糖-2-羟基松属素；
(B)CGTb催化C-葡萄糖-2-羟基柚皮素生成C-***糖-C-葡萄糖-2-羟基柚皮素，CGTb催化C-葡萄糖-2-羟基松属素生成C-***糖-C-葡萄糖-2-羟基松属素。
具体实施方式
下述实施例中的实验方法，如无特殊说明，均为常规方法。下述实施例中所用的实验材料，如无特殊说明，均为自常规生化试剂公司购买得到。
实施例1：黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中C-糖基转移酶的克隆、原核表达、酶蛋白纯化和功能表征
1.黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中C-糖基转移酶基因的筛选
从NCBI中下载黄芩的转录组数据(SRR3123399-RNA、SRR3130396-RNA、SRR3727161-RNA、SRR367956-根、SRR3367955-叶)、甘草转录组数据(DRR006519、DRR006520、DRR006521、DRR006522、DRR006525、DRR006526、SRR1783599、SRR1783600、SRR1783602、SRR1811619、SRR2537378)、少根紫萍转录组数据(SRR647050、SRR647053、SRR948161、SRR2045955、SRR2045956、SRR2045957、SRR2045958、SRR2048534、SRR2048535、SRR2048536、SRR2048537、SRR2048538)。水稻与玉米使用在线转录组数据库。大薸、一把伞南星进行转录组测序。
对于NCBI上下载的转录组数据，用SOAPdenovo-Trans-src-v1.04程序进行转录组数据的拼接。将拼接得到的序列文件，与已知的植物的碳苷转移酶序列(OsCGT、UGT708A6、FeCGTa、FeCGTb、UGT708D1、MiCGT、FcCGT、CuCGT)进行本地BLAST比对，选择评价参数小于10-21的序列分析其ORF。例如在黄芩中一共得到11条完整的ORF序列，其中SbCGTa和SbCGTb含有植物黄酮碳苷糖基转移酶的保守区(DPF)，且与大部分植物C-糖基转移酶氨基酸序列相似，通过系统进化分析，聚在一类。因此，这两条序列很有可能为黄芩C-糖基转移酶序列。
基本保守区进行候选基因的筛选。甘草中一共拿到3条候选碳糖基转移酶基因；一把伞南星中一共拿到2大类含有DPF保守区的候选基因，每类中均有多条相似度较高的基因；少根紫萍中一共拿到2条含有DPF保守区候选基因；大薸中一共拿到5条含有DPF保守区候选基因；玉米中一共拿到5条候选碳糖基转移酶基因；水稻中一共拿到3条候选碳糖基转移酶基因。
2.黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中C-糖基转移酶基因克隆、原核表达载体构建、转化入大肠杆菌、诱导蛋白表达
选取植物幼苗，按照TranZol法(ET101,Transgen Biotech,China)提取RNA。按照SMARTRACE cDNA Amplification Kit法合成cDNA第一链(Clontech Laboratories,Inc.)。设计引物用于ORF的扩增，引物序列如表1所示，以cDNA为模板，扩增ORF，PCR条件：94℃预变性3min，94℃变性30s，56℃退火30s，68℃延伸1min 30s，36个循环，酶：TransStart KDPlus DNA Polymerase(Transgen Biotech,China)。回收ORF，检测DNA的浓度应大于50ng/μL。
表1.引物序列表
黄芩和水稻C-糖基转移酶重组载体(pET28a-SbCGT/OsCGT)的构建采用Quick-change法，PCR条件：98℃预变性3min，98℃变性20s，60℃退火30s，72℃延伸4min，36个循环，酶：Q5High-fidelity DNA polymerase。其中PCR产物的含量≥250ng，体积小于5μL，pET28a空质粒的含量在30～50ng之间，PCR反应体积为25μL。反应结束后，取20μL进行DpnI消化处理，加入DpnI 2μL,CutSmart 3μL，ddH2O 5μL,37℃酶切5个小时或过夜酶切。将酶切后的溶液加入到200μL的DH5α感受态细胞中，进行转化。涂布于含卡那霉素的LB平板上，37℃过夜培养，选取单菌落，PCR筛选阳性转化子(采用pET28a通用引物)。
玉米、甘草、大薸、一把伞南星采用同源重组的方法构建pET28a(+)重组表达载体，即目的片段两端和酶切后的pET28a(+)线性载体两端分别具有20-25bp左右的同源序列，进而通过无缝拼接试剂盒(Transgen)进行同源重组。采用BamHI进行pET28a(+)载体单酶切，37℃酶切2h后进行电泳以及胶回收，载体浓度大于100ng/μL。同源重组体系：ORF(100ng/μL)0.5μL，酶切后载体(100ng/μL)2μL，Assembly Mix 5μL，水2.5μL。50℃反应15min后导入Trans1-T1感受态细胞。涂布于含卡那霉素的LB平板上，37℃过夜培养，选取单菌落，PCR筛选阳性转化子(采用pET28a通用引物)。
将得到的阳性单菌落在新的含卡那霉素的LB平板上划线保存，并挑选菌落摇3mL，提取质粒，测序，选取阳性质粒，导入到BL21(DE3)感受态细胞中，涂布于含卡那霉素的LB平板上，37℃过夜培养，选取单菌落，PCR筛选阳性转化子(采用pET28a通用引物)。将得到的阳性单菌落在新的含卡那霉素的LB平板上划线保存，并挑选菌落摇3mL(含卡那霉素)，37℃过夜培养，放大至500mL(含卡那霉素)，培养至OD600≈0.6，加入0.1mmol/L的IPTG，19℃诱导20h，7000rpm×5min收集菌体。
3.黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中C-糖基转移酶蛋白的纯化
在菌体沉淀中加入裂解缓冲液15mL，溶菌酶10mg，冰上放置30min；冰上超声(200W,10s,10s,10min)，10,000rpm 4℃离心30min，取上清；加0.35mL 50％Ni-NTA，轻轻混匀，200rpm 4℃磁力搅拌60min后，Lysate-Ni-NTA上柱子(柱子先用lysis buffer冲洗)；Flow-through流过，收集流分FT；清洗缓冲液冲洗两次，每次5mL，收集流分W-1，W-2；用不同咪唑浓度的洗脱缓冲液冲洗(50mM×3、100mM×3、150mM×3、200mM×3、250mM×3)，收集流分；取100μL Bradford加入1μL流分，检测蛋白浓度确定上样量；一定上样量的流分加洗脱缓冲液补足至20μL；加5μL上样，煮沸5min，冷却后短暂离心，4℃保存；SDS-PAGE。
蛋白的纯化保存(超滤离心法15mL/30KD)：洗脱缓冲液冲洗滤膜，4℃，4000rpm×5min；合并流分(约4.5mL)，4℃，4000rpm×12min，剩余0.5mL；加存储缓冲液3mL，4℃，4000rpm×30min，剩余0.5mL；加存储缓冲液3mL，4℃，4000rpm×20min，剩余0.8mL；分装100μL×8，-80℃保存。
蛋白浓度的测定：按照BCA蛋白浓度测定试剂盒说明书测定蛋白浓度，使用的蛋白标准为牛血清白蛋白。
4.黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中C-糖基转移酶的功能表征
黄芩中C-糖基转移酶SbCGTa和SbCGTb的功能表征：参见图2至图4，SbCGTa催化反应体系包括100μL的磷酸盐缓冲溶液(pH 6.0)，底物(0.2mmol/L)：2-羟基柚皮素(2)、2-羟基松属素(1)、根皮素(7)、夫洛丙酮(10)和2,4,6-三羟基苯甲酸甲酯(11)，UDPG(0.25mmol/L)，适量的SbCGTa。SbCGTb催化反应体系包括100μL的磷酸盐缓冲溶液(pH 6.0)，底物(0.2mmol/L)：C-葡萄糖-2-羟基柚皮素(4)、C-葡萄糖-2-羟基松属素(3)、3’-C-葡萄糖根皮素(8)、3-C-葡萄糖夫洛丙酮(12)和2,4,6-三羟基-3-C-葡萄糖苯甲酸甲酯(13)，UDP-***糖(0.25mmol/L)，适量的SbCGTb。30℃反应1h，加入400μL甲醇终止反应，真空旋蒸干燥后，加入100μL 50％的甲醇复溶，经UPLC/MS检测。
如图2所示，SbCGTa可催化2-羟基柚皮素(2)和2-羟基松属素(1)，分别生成C-葡萄糖-2-羟基柚皮素(4)和C-葡萄糖-2-羟基松属素(3)；而SbCGTb可催化C-葡萄糖-2-羟基柚皮素(4)和C-葡萄糖-2-羟基松属素(3)，分别生成C-***糖-C-葡萄糖-2-羟基柚皮素(6)和C-***糖-C-葡萄糖-2-羟基松属素(5)；C-***糖-C-葡萄糖-2-羟基柚皮素(6)经HCl处理脱水生成夏佛塔苷(6b)和异夏佛塔苷(6a)，C-***糖-C-葡萄糖-2-羟基松属素(5)经HCl处理脱水生成白杨素-6-C-α-L-***糖-8-C-β-D-葡萄糖(5a)和白杨素-6-C-β-D-葡萄糖-8-C-α-L-***糖(5b)。
SbCGTa可催化根皮素(7)生成3’-C-葡萄糖根皮素(8)，而SbCGTb可催化3’-C-葡萄糖根皮素(8)，生成3’-C-葡萄糖-5’-C-***糖根皮素(9)，其反应式和检测结果如图3所示。
SbCGTa可催化夫洛丙酮(10)生成3’-C-葡萄糖夫洛丙酮(12)，而SbCGTb可催化3’-C-葡萄糖夫洛丙酮(12)生成3’-C-葡萄糖-5’-C-***糖夫洛丙酮(14)(见图4)。SbCGTa可催化2,4,6-三羟基苯甲酸甲酯(11)生成2,4,6-三羟基-3-C-葡萄糖苯甲酸甲酯(13)，而SbCGTb可催化2,4,6-三羟基-3-C-葡萄糖苯甲酸甲酯(13)生成2,4,6-三羟基-3-C-葡萄糖-5-C-***糖苯甲酸甲酯(15)(见图4)。
玉米、水稻、甘草、大薸、少根紫萍、一把伞南星的功能表征与黄芩类似，结果如图5所示。
以上结果表明，SbCGTa、GuCGTa、AeCGTa、LpCGTa、PsCGTa为2-羟基黄烷酮C-糖基转移酶，而ZmCGTb、OsCGTb1、OsCGTb2、SbCGTb、GuCGTb、AeCGTb、LpCGTb、PsCGTb为C-葡萄糖-2-羟基黄烷酮C-糖基转移酶基因。这些CGTa和CGTb在黄芩、玉米、水稻、甘草、大薸、少根紫萍、一把伞南星中按照先后顺序、依次催化合成夏佛塔苷和异夏佛塔苷。这些酶可以用于体外酶法合成夏佛塔苷、异夏佛塔苷、白杨素-6-C-α-L-***糖-8-C-β-D-葡萄糖、白杨素-6-C-β-D-葡萄糖-8-C-α-L-***糖、3’-C-葡萄糖-5’-C-***糖根皮素、3’-C-葡萄糖-5’-C-***糖夫洛丙酮和2,4,6-三羟基-3-C-葡萄糖-5-C-***糖苯甲酸甲酯。
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SEQUENCE LISTING
<110> 北京大学
<120> C-糖基转移酶及其在合成夏佛塔苷和异夏佛塔苷中的应用
<130> WX2019-BY-003
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<170> PatentIn version 3.5
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cagatcgacg ttgcaaatcg cgacgccatt gagatccccg atttaggccc aatcagtatg 540
tccagtattc cccccaaaat gctggacccg agcgatttct tctccgcctt catcagttca 600
aacgtttcgt ctctacataa agtcaaaggg gttctcatca acacgttcaa tagcttcgaa 660
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ctccggtggg aggccctgcg ccgctccttg cacctcctcg gcccggtcat cgccggcgtc 360
accccgcgcg tcacggccac cgtcaccgac gtcacgctgg tgtcccacgt caaccccatc 420
gccaaggacc tgggcgtcca gtgccacgtg ctgtacgtct cctccgcggc catgatgtcc 480
ctctgctcct acttccctat ctacctcgac aacaaggatg ccggggccga cgtcggcgac 540
gtcgacatcc ccggtgtgcg tcgcctcaag aggtcctggc tgccgcagcc attgctcgac 600
ctgaacaagc tcttcacgaa gcagttcatc gagaacggcc gggagatggt caagaccgac 660
ggcgttctga tcaacacgtt cgacgcgctg gagccggtgg cgctcgccgc cctccgcgac 720
ggcaaggtcg tccgcgggtt cccgccggtg ttcgccgtcg gcccgcacag ctccctggcg 780
agcgaggcga cgaagggcgc cgccgccgac gcagaggggt cgccgatggc atggctccgc 840
cagcagccgg cgcggtcggt ggtgtacgtc gccttcggca gccggtgcgc cgtgagccac 900
gagcagatcc gggagatcgc ggcggggctg gaggcgagcg gctcccgctt cctgtggatc 960
ctcaagacca cggtggtgga ccgcgacgac gacgccggca tccgcgacgt gctcggcgac 1020
gggttcctgg agcgcgtgcg ggggcgcggc gtggtgacca aggcgtgggt ggaccaggac 1080
gcggtgctca gggacccggc ggtgggcctg ttcctgagcc acagcgggtg gaactcggtg 1140
atcgaggcgg cgacggccgg cgtgccgctg ctggcgtggc cgcggggcgg cgaccaccgc 1200
gtggcggcga cggtggtggc gagcagcggc gtcggggtgt ggatggagca gtggagctgg 1260
gacggggagg agtgggtggt gagcggggag gagatagggg ggaaggtgaa ggagatgatg 1320
gccgacgccg gcgtgcggga gaaggcggcc aaggtcggcg aggaggtggc caaggcggtg 1380
gccgtcggcg gcacgagcca cacgggcatc ctggatttcg tcgccaagct caaggctgca 1440
acttaa 1446
<210> 5
<211> 1419
<212> DNA
<213> 甘草 (Glycyrrhiza uralensis）
<400> 5
atgggggaaa acaacatgtc ctccattgct cctcaccctg ttgttcatgt ggctctcttg 60
ccaagtgcag gaatgggaca tctctcccca tttctaaggc tagcctctct gctcctccac 120
caacactgcc acgtcaccct catcacccct caactcacag tatccaaagc agaagaggat 180
cttctctcaa ggttcctctc agcattccct caagtcaacc aactccactt ccatctcccc 240
ccttctgatt ccactatttc cactgaccct ttcttccttc aatttgcttc cattcgtagc 300
tcttcccacc ttctcacccc tcttctgtct tcactcacac cccctttgtc ttcattcatc 360
tatgacatga cccttatttc accacttctt cccattgctg aatcccttgg tgtccctcac 420
tacatcctct tcacttcttc agctaccatg ttctctttct tctcttactt cccaacactg 480
gctaaatctg aatcctttcc tggaaaactt gattttgttg agattcctgg ggtttcagtt 540
tcatccatac ccagatcatc aatcccacca ccacttttgg tcccaaacag tctctttggc 600
aaactcttca tggaggatag tcctaagctc aagaaacttc atggggtttt ggtcaacact 660
tttgaaggga tagaaaaatt gtcccttgag gctctcaacg gtgggaaagt ggtaaaaggg 720
ttgcccccag tttatggggt tggtcccttt gtgccttgtg agtttgagaa ggttgtgaaa 780
agaggggaaa caatatcgga gtggcttgat gaacagccta gtggttctgt tgtgtatgtt 840
agctttggga gtaggacagc aatggggagg gaacaactaa gagaggtagg tgatgggttg 900
gttaaaagtg ggtggaggtt tttgtgggtg gtgaaggata agatagttga tagggcagaa 960
gaagaagggc tagatggggt gttggggttt gaacttgtgg agaggatggt aaaggagaag 1020
aagggtttgg tggtgaagga atgggtggat caaagtgaga ttttgggaca caaagctgtt 1080
ggggggtttg tgagtcattg tgggtggaac tcagtggtag aggcagcatg gtttggtgtg 1140
aaaatcctgg ggtggcctct gcatggggat cagaagataa atgctgaagt ggtggcaaag 1200
ggagggtggg gagtgtggaa agaagggtgg gattgggaag gggaacgttt ggtgaaaggg 1260
gaggaaattg gagaggctat aagagaggtg atgaatgatg agtctcttgt gatgaaagca 1320
acacaggtga aaaaggatgc aaggaaggcc atcagtgttg gtgggggatg tgaggttgct 1380
cttcagaaac taatggaagt ttggaagaag aatgtttga 1419
<210> 6
<211> 1458
<212> DNA
<213> 甘草 (Glycyrrhiza uralensis）
<400> 6
atgacttcct gtactaattc agaacccgtc accgtccacg ttgctttcct tccaagtgca 60
ggcatgggac atctcaaccc atgtctcaga acagccgaac agttcctccg tcacggttgc 120
aaagtcactc tcatcactcc caaacccacc gtctcactcg ccgaatcaaa cctcatcaac 180
cgtttctgtt cttctttccc ttcttcacaa ctcaaccagg tagacttaaa cctcatcgat 240
gtggacccta cacaagtcaa caccaatgac ccattttggc ttcacttcga aaccattcgt 300
caatcccttc aataccattt acccacaatc ctatcctcac tctcatcacc accactctct 360
gctttcatct atgatgtgag tttgatttcc cctttggtac ccatcactga aaaactctgt 420
tttcctagct acatttactt cattgcacct gctagaatgt tctctttttt tgcacacctc 480
tctgttctct ctgagaaaaa cccaggtgtg aaaccttctt ctttcatcgg tggtgttgtt 540
gagattccgg gtattgctcc gatacccaga tcctctgttc ctcctctgct tcttgagcct 600
aatagtttgt ttgagagtat atttatggag gatagtccta aactgaagaa gcttagtggg 660
attttcatca actcgtttga ggaacttgaa ccggaggcac ttgctgcact caacaatgga 720
agggttgttc ctgggttgcc tcctgtgtat cccattggtc ccttggtgcc ttgtgagttt 780
gagaaggtac ggtcctcacc caagactaac tcgagtacta tattgaagtg gcttgatgga 840
cagcctgcag ggtcagtggt atatgtttgc ttaggaaaca ggacacaaac aagaagggag 900
caaataaagg acatggctca agggttaatt tcttgtgggt acaagttttt atgggtggtg 960
aagctgaaga tagtggacag agaagatcaa gaggaagagt tggatgatgt gttggggaat 1020
gaggtgatga agcaggtgaa tgaaaagggt atggctgtga agacatgggt tgaccaaatg 1080
gagattttgg gtcacccttc agttggaggt tttgtgaacc atggtgggtg gaactcattg 1140
gtggaaggtg tgtggcatgg gaagcctatt ttatcatggg cacatgatgg ggatcagaaa 1200
atagcctcag aggttgtgaa gatgaatggg gtggggattt ggccacaggt gtggggatgg 1260
ggagcaaggg atgatgtggt gaaaggggag gagattgcaa gggcaatcaa tgagatgatg 1320
agcaatgagt ccttgaggaa caaagcaaga gaaatgaagg aggctgctag gaaggctgct 1380
ggtgttggtg ggagttgtga ggttattatt aagagacaaa ttgaggaatg gaagaagagg 1440
attgttcaag gcatctga 1458
<210> 7
<211> 1422
<212> DNA
<213> 一把伞南星 (Arisaema erubescens)
<400> 7
atgtcgccgc cgacggctga gggggctcgg acaccccgag tggccctcct ccccagcgcc 60
ggcatgggcc atctcacccc cttcctccgc cttgccgccg tcctggccgc ccgcggctgc 120
gacgtcaccc tcatcacccc ccatccggtg gtttccgctg ccgaggcccg ccacatcgcc 180
gccttcctcg cgtccgcgtc cccccgggtc cgctccctcg agttccgcct cctccccttc 240
gacgccgcct ccgccacctc caccgacccc ttcttcctcc agttcgaggc catccgccgc 300
tccgcccacc tccttggccc cctcctcgct tccgcctgcc cggccctatc cgctgtcgtc 360
accgacgtca gcctcatctc cgccgttgcc cctgtcgtcg ccgccgtcgg ccttcccaac 420
tatgtcattt ttacttcctc ggccgcgatg ctcgcgctct gcgtcgcctt cccctccgcc 480
tcagcatccc tcggcggcgc tgccgacacc ttcgaggtcc ccgggttggc cgcgctgccc 540
atcaactggc taccgcagcc ccttcgcgac ccggggaacc tgttcacgac ccagttcgtg 600
gagaacggcc gggaactcac ccgagtcgac ggcatcatga tcaacacctg ggacgcgctg 660
gagggcgaga ctctcgccgc cctcaacgct ggggcggtgg ctcatctccc cccggtaatc 720
cccgtcggcc ctctcccgcc ggtgaaaccc cgcagcatcc ccggcatccc ctggctcgac 780
gaccaaccgg agcgctctgt cgtctacgtc agtttcggga gccggacggc gctgtcggcg 840
gagcagatca gggaactagg cgctggcctg gagaagagcg ggtgccggtt cctgtgggcg 900
gcgaagagca agaaggtgga taaggaggac gacggggtgg tgctggagga gctgctagga 960
gacgacgggt tcctgcgaag gatggagggg aggggcctgg tggtaaagga ctgggtcgac 1020
caggacgccg ttctggcgca ccctgccgtg gggggcttcc tctgccactg cgggtggaac 1080
tcggtgacgg aggcggccat tcacggggtg cgcgtgctgg catggccccg gcacggcgac 1140
cagcggctta acgcgtgggt ggtggagagg ggcgggctgg ggacgtggcc gacagagtgg 1200
agctgggaag gggacgccga gctggtgagc ggggaggaga tcgctcggcg ggtgagggag 1260
ctcgtgggga gccccgacgc cagcgacgcg gcggcaccag tgggcgagga ggccgtgaga 1320
gccgccgagg cctgcggcag ctcccgcaag gccctggacg agctcatcga gaagtggggc 1380
cggcaactgg ggacgaggtc agatggtatg cgtcggccat ag 1422
<210> 8
<211> 1458
<212> DNA
<213> 一把伞南星 (Arisaema erubescens)
<400> 8
atgtctcggg aacagcagtc gtcgagctcg aacggtggcc tcacagtgcc ccatgtcgcc 60
gtgatcccca gcgccggcat gggccacctt tcgatctccg tccgtctagc ggcgaccctg 120
gcctcccacg gctgcctggt taccttcata accccgcggc ctgtcgtgtc cgacgcggag 180
tcccgcacca tcgcggatct ccattcctca tatcctcaga tccggtccct cgaccttcgc 240
ctcctcccct tcgacgcccc cgatgtcgac cccttccttc ggcagtttga agtcattcgc 300
cggtcggcaa gcttgctgca acccctcctc acctctgtta caccccctct ctctgccgtc 360
atcgttgaca tcagcctgct ctccaccttt gtccccgtca ccaaagccct ccgcctcccc 420
acgtatgtga ccttcatctc ctgcgccatg atgctcgccc tgctggccgt cttccccagc 480
gtcaggaaag cagcaggtga cggaccaatc gccgccgtgg acattccggg cgtagggacg 540
tcgctgcctg gttcgtggat cccaccgccc ctacacgatc cgaagcatct cttcgcgatc 600
cagttcctgg agaacggcga ggcgctgctg caggcggacg gcgtctttat caacaccttc 660
tccgccctag aggcagaaac tctggccgcg ctcaacgccg gcaacgtcgt gccgggcctc 720
cctcccgtca tcgccatcgg tccacagcaa cccgtcaagt tcgagggagc caagcgcgct 780
gcagcggccg ggagggacga gtgggcggcg tggctggacg cccagccacc tgggtcggtg 840
ctgtacgtag ccttcgggaa caggacggcc ctgccgaggg agcagttaag ggagctgggc 900
ttgggcctgg agaagagcgg ctgtagattc ctgtgggtgg tgaagagcaa gattgttgat 960
agggaggacg acgctggtgt gttgttggag gacctgctgg gagaaggata ccgggagaga 1020
gtgggggaga gagggatggt ggtgaagagc tgggtggacc aggaagaggt cctcgcccat 1080
ccctccgtcg gcgggttcct cagccacgcc ggctggaact ccgtggcaga ggcggcgctg 1140
cagggcatct gcctttttgg ttggccggcc ggcggcgacc agcgcgtggc tgccttggta 1200
gtggagaagt gcggtgcggg ggcgtgggtg aaggagtggg gctggtccgg cagcgggcgc 1260
ggccttgagg agacgccgac ggtgagtgga gaggagatcg ggaagcgtct gagggagctg 1320
atgggggacg cgtccctaag ggagtccaca gcgcgcatcc gtaaagacgc cctaaggtcg 1380
gtcggcgtcg gcgggaagtc ctacgaggcg gtgcggagct tcatcgccgg actgggcggc 1440
gtgggataca ccacatga 1458
<210> 9
<211> 1455
<212> DNA
<213> 少根紫萍 (Landoltia punctate)
<400> 9
atgagaccgg ggagctcgga gaggaatcat cccgcagagc gacccacaac aatcctctcc 60
gggagtatgg caccggcggc ggttcctcac gttgccctgc tccccagctc cggcatgggg 120
cacctcaccc cgtttcttcg tctcgctgct gcgctggcct cgcacggctg cgtcataacc 180
ttcattactc cgacgccggt ggtatccgcc gccgaggctc gccatgtcga ggccttcatc 240
tcttcttctc cgctgttccg gcggctcgag tttcccctcc tcccttttga cgtctcctcc 300
gtcgtctccg acgacccctt cttcctccag ttcgagagga tcagccgatc cgcgcgccac 360
ctcggccccg tcctgtcgtc tgtgtctccg cctctgtctg ccctgatttt ggatgtaacc 420
ctcacctcca gcatactccc cattgccgcc gaaatctctc tgcccgctta cattctcttt 480
acctcatcgg cgggcatgct ctccctctgc ctctcctacc cggagattgc cgcctccgga 540
ggcgtgacca tcaagatccc cggcgtggcg gaggacctgg cacccagctc cctgccccag 600
ccgttacgag acccgaggaa cctcttcacg ggccagttca tcgagaacgg acgagcaatg 660
gcgcgagcgg acgggatcat catcaacacc tgggaagccc tcgagccggc gactcttgcg 720
gcgctccagg gcagtaaggc ggtatctggt tttccattgg tgatccccgt cggtcccctc 780
cttgccgcaa gcgacatcca cacagccgcc gacgacttgg tcatcccctg gctcgacgcg 840
cagcctgctt cctcggtcgt ctttgtcagc ttcggcagtc gcacggcgct gtcggcggaa 900
cagctgcggg agcttgcagc cggactggag agcagcgggt gccggttcct gtgggtcctg 960
aagaccaaga aggtagacag ggaggaccaa gagggcgaga aagccgtgga tgagctgctc 1020
ggagagggct ttctgcaaag ggtggagtgg aaggggaagg tggtgagcgg gtgggtggat 1080
cagcgggctg ttctggacca ccccagcgtg ggcggcttcg tcagtcactg cggctggaac 1140
tcagttacgg aggcagcgct aggagggatg agggtgctgg cctggccgcg tcacggcgac 1200
cagaggatca acgccatggt agtagagaag agtgggctgg ggaagtggcc cagcttatgg 1260
acgtgggaag gagacgacga gatagtgcgg agagaggaga tcgccgggag agttgcagag 1320
cttatggcta gccccgccgc ggcggccgcc gccgccaagg taaaggagga ggccgtcagg 1380
gccgcgaccg cgggcggcag ctcgcaacgc caactggagg acctcgtctc gcggttcacg 1440
tgttcgggag attga 1455
<210> 10
<211> 1407
<212> DNA
<213> 少根紫萍 (Landoltia punctate)
<400> 10
atgtctcctc ctgcgcctgc ggacgtagtc tcgagtgcaa aaccgcacgt agctgtcatc 60
ccggccgcgg gcatggggca cctcaacccc accctgcgcc tcgccggcga gctggcctcg 120
cgcggttgcg tcgtcacctt catcaaccct tcgccgccgg tgagcctagc ggaggctacc 180
tccgtggcag agttcgtcgc ctccacgccc ggcgttcgcc tgctggactt gcccgtccag 240
cccttagacc cctcctgctt ccccgcccac gaagatccgt tcctccgtca gttcgaggcc 300
gtccgtcgtt ctgcgcctct gcttaccccg ctcctctccg acgtatcccc tccactcgcc 360
gccatcgtct gcgacatcgc catctgctcc accttcctaa ccgttgctgc cgagatttcc 420
ctccccgcct acgtcttctt ctccctttcg gcgcagatgc tatccctcaa cctggccttc 480
cccaccgtcg ccgaccaggt ctacggtgct ggcgaaggcg acgagatacg cttccccggc 540
ttgccggaga gcatacccag gagctggctc cctccgcccc ttctcgaccc cgcccaccta 600
ttcgctgtcc acttcgttga gaacgggaag gcaatgccgc gagcggcagg catcctggtc 660
cactcctggg aagcgctgga gccggaggcc ctggcggcgc taaggggcgg cagagtgctc 720
gccggcctgc cgccggtgct gcccatcggc cctctgtacc agaaagagaa gtcgaacgcc 780
gtgttcttgc cttggctcga cgctcagcgg gatcgctcgg tgctgtttgt ctgcttcggc 840
aaccgctcga cgcattcgcc ggagcagctg cgggagatgg ccgccggctt agagaggagc 900
ggctgccggt tcgtgtgggt gctgaagacg aaggtggtgg acaaggacga ggacgaaggg 960
gcgcagaagg agatactcgg agagggttac ctcgaaagag ttaaagagag gggggttgtg 1020
atcaacgggt gggtcgacca gatgaccatc ctgtctcacc gcgccgtagg cggcttcttc 1080
tcccacagcg gatcgtcctc ggtggccgag gcggctattg gagggcagcc gcttctgctg 1140
tggcctatgg gaggcgatca aaggatgtcg gccctagtgg cggagaggag gggtatgggc 1200
gtgtggccga gagggtgggg ctggtctgct gatgacaagc tgattccagg ggaggagatc 1260
gcccggcgga ttaaggattt catgggggac aacgccttgc gagccgtggc ggccaagatg 1320
aagaaggaga cggcaagcgc catggccccc ggcggcagca aggatcagtg gttcgatgac 1380
tttatcgcga ggatcaaccg cgtctga 1407
<210> 11
<211> 1485
<212> DNA
<213> 大薸 (Pistia stratiotes)
<400> 11
atggttcaag agacatcacc gccggtgtgc aattttaggg acactgtcag cacgacatcg 60
ccagctccac acatcgccat cctttccagc gccggctttg gccacctcat tgtctccgtc 120
cgcctggcgg tggccctcgc ctcacacggt tgcctgatca ccctcgtcac gccgctcccc 180
acggtgtccg aagccgagtc ccgctacatc tccgacttcc actcctgctt cccccacatc 240
aggctactcg agttccacct cctccccttg gacgcccaag cagacaccga ccccttcttc 300
agccagttcg aaaccatccg ccgatctgct gccttgctgg ggcctctcct tgcctcggcc 360
acgcccccgc tctccgctct ccttgccgac attagtgtgc tctcttccta catccccgtc 420
acgaaagccc tcggcatccc cacctacgtg actttcacct cctgcgcggc catgcttgcg 480
ctgtcttgcg ttttcccgag agtcagaaaa gaggtggggg acggtccggt cgtgtctatt 540
gacattcccg ggctggggac cgtgccgggt tcgtggattc cgccgcctct gcacaacccc 600
aaccatatct tcgcgaggca cttctcggag aacggggcgg cggtgctgca ggcggacggc 660
gtccttgtga acacgttcga agccctggaa ggagatattc tgtccgcact taacgctggc 720
actgttttgc cgggcctccc acccgtcatc gccatcgggc cacaggaacc tcttaagctt 780
ggagcgtcgg cgcggccggc aggggaacca gacggttggg cgacatggtt ggacgcccag 840
cctcccaagt cagtgctgta tgtagccttc gggaacagaa cggccctatc cactgtgcaa 900
atacgtgagc tgggcaaagg tctggagaag agcggctgca ggttcctgtg ggtggtgaaa 960
agcaagatcg tggacaagga cgagagcgtg gagttggatg agcttctggg cgaaggatac 1020
ctggagagag taggagacag agggatggtg gcgaagagct gggtggatca ggaggaagtt 1080
ctcgcccacc cctcagtcgg cgggttcctc agccacggcg gttgcagctc ctcgcaggaa 1140
gccattctgc acggggtacg gtcgctgaac tggccgggca tgggcgacca gcggatggtc 1200
gccgcacagg ccgaaaagcg cggggctggc gtttgggtga aggagtgggg ctggagtagt 1260
cgcgtgcacc gggaggagcc agaaccggtg agcagcgagg agatcgcgaa gcgtgtgaag 1320
gagctgatgg aggacgacgc ggtgggggag gccatgcgac gtatccacga ggaggccgta 1380
atggggctcc ggcccggcgg cagctcctac gaggggctgc gcaagttcgt ggccctcgta 1440
gaggcgaaga gacgatcatc accaccgcag gtttcagaga cctaa 1485
<210> 12
<211> 1452
<212> DNA
<213> 大薸 (Pistia stratiotes)
<400> 12
atggcccgag agccatccca gctggtgcgc gatggcgacc tgatcccaac gccaacccct 60
cacatcgtca tccttcccag cgccggcatg ggccacctca cggtttccac ccgcctggcg 120
gcggccctcg tctcacacgg ttgcttggtc accctcatca caccgctccc catggtgtct 180
gaagccgagt ccaactgcat ctccaatctc tgctcctcct tcccgcacat caggttgctc 240
gagctgcatc tcctcccctt cgacgccccc gacgtcgacc ctttcctccg ccagttcgag 300
tccatccgac gctcggcggc cttgctggag cctctcctcg tctctgttac tcccccagtc 360
tccgctctcc tcaccgacgt caccgtgttc tcttccgtcg tccccgtcac caaagccctc 420
cgcatcccta tctacatgat cttcaccacc tgcgcctcca tgctggcact ggttagcact 480
tttcccggca tcaggaaaga ggcaggggac ggcccgatcg aatctataga cattcccggc 540
gtcgggacga taccgggttc gcggattccg cggcctctgc acgaccccaa acacctcttc 600
acgatccagt tcgtggagac tggagaggca atgctgcagg cggatgggat cttgtcaaat 660
accttcgaag cccttgacgg ggatattttg gcggcgttaa acgctggcgc cgtcgtgccg 720
ggtttccctc ccgtcatcgc cgtcgggccg caggagccca tcaaactcag agcggccgcc 780
cgaccggcag tagaacgaga cgactggatg gcatggctgg acgcccagcc gcccaaatct 840
gtgctgtatg tagcctttgg gaacagaacg gcgcagtcga gggagcaaat aaaggagctg 900
ggcataggtc tggagaagag cggctgcagg ttcctgtggg tggtgaaaag caagattgtc 960
gacaaggacg acgacgatgg cgtggagctg ggtgagctgc tgggtgaagg gtacctcgag 1020
agagtgggag agaggggagt ggtggtgaag agctgggtaa tccaggagga ggtcctcgcc 1080
cacccctcgg tcggcgggta cctcagccac ggtggaggga actccacgtt cgaggccgtg 1140
ctctatgggg tgcccccgct gatctggccc ggaggcggcg accaacagat ggtggccgcg 1200
caggtggaga agcgaggggt gggcgtgtgg ataaaggagt ggggactgtg cggcgtgaac 1260
caggtgagca gcgacgacgt cgcgaagcgt gtgagagaaa tgatggaggg cgacaatgcc 1320
gtcggcgaat ccgtgcgacg gatgcaagag gaggccgtga ggggactcgg gcccggcggc 1380
agctcgtacg agggactgcg caagttcatt gccggcgtag tggagagggg gtcatcacca 1440
ccggaaatct aa 1452
<210> 13
<211> 1452
<212> DNA
<213> 玉米 (Zea mays)
<400> 13
atggccccgc cggcaacgaa tagcccggga gaggtcgaca gcggcggccg cccgcatgtc 60
atgttcatcc caagcgctgg catgggccac ctcatcccat tcttccgctt catcgcggct 120
ctcgctgggc agagcgtcga catctctgtg gtgactgtcc ttcctaccgt ttctgccgcc 180
gaggaggact acttcaccag cctcttcgct gccctccccc gtgtccgccg cgtcgacctc 240
cacctcctgc cgttcgacgc gtccaagctc ccaagcaacg accgcgaccc gttcgttctc 300
cggtgggagg ccctgcgccg ctcagcgcac ctcctgggcc cgcttattgc cggcgccgcc 360
ccacgcgtct cggccgtcat tacagacgtg accctgacct cccacgtcat ccccatcgcc 420
aaggagctcg gcgtccagtg ccacgtcctc ttcccctcca gcgccaccat gctctcgctc 480
cttgcttaca cccctgtaca catcgacaag aaggccgagc aggggagtga catcggcgac 540
gtcgacatcc ccggtgtgtg tcgcataccg cagtcttgtc tcccgcaggt gctgcttcac 600
ctcgacaaac tcttcaccaa gcagttcatc gccaacggcc gcgagatcat caacgctgac 660
ggctttctgg tgaacacgtt cgacgccttg gagccggtgg cgctcgctgc cctgagagat 720
ggcaaggtcg tcccggggtt cccaccggtg tacgccattg gcccactcat gtcgcagcaa 780
aactcagttg acgaaggtgg gaaggaagaa ggatcgggct cccccgttgc ctggcttgac 840
gagcagccag cgcggtctgt ggtgtacgtc gcgttcggca accgctgcgc cgtgagccat 900
gaccagatca gggagatcgc cgccgggctc gaggcgagca actgtcggtt cctttgggtg 960
ctcaaaacca caacggtgga tagggacgac agcgccgtgc tgacggaatt gctcggcgag 1020
gagttcctgg agcgagtgca gggtcgcggc ctcgtgacca aggcgtgggt ggaccaggag 1080
gccctgctga agcacccggc catggggctg tttctaagcc acagcgggtg gaactcggtg 1140
accgaggcgg ctgccgccag cgtgccgctg ctggcgtggc cccgcggcgg cgaccaccgc 1200
gtgaacgcca tggtaacggt gagcggtggg gttgggatgt ggatggagca ctggagctgg 1260
gacggggagg actggctggt gaccggggtg gagatcgcga agaaagtaaa agaggtcatg 1320
tccgacgaag cggtcagggc aaggacgacg aggaccgccg aggaagcggc caaggcagtt 1380
gcagagggtg gcaccagcta ccggagcatg gaggagttta tttccagcgt caaagcaacc 1440
gtggcgtctt ga 1452
<210> 14
<211> 459
<212> PRT
<213> 黄芩 (Scutellaria baicalensis)
<400> 14
Met Ala Ser Thr Thr Lys Ser Glu Asn Val Gly Ala His Ile Ala Leu
1 5 10 15
Phe Pro Cys Ala Gly Met Gly His Leu Leu Pro Phe Leu Arg Leu Ala
20 25 30
Ala Met Leu Asp Ala Arg Gly Cys Ala Val Thr Val Ile Thr Val Lys
35 40 45
Pro Thr Val Ser Ala Ala Glu Ser Asp His Leu Ser Ala Phe Phe Thr
50 55 60
Ile His Pro Arg Ile Thr Arg Leu Glu Phe Gln Leu Leu Pro Tyr Gln
65 70 75 80
Lys Ser Gly Leu Arg Asn Asp Asp Pro Phe Phe Ile Gln Met Glu Thr
85 90 95
Ile Ala Thr Ser Val His Leu Leu Arg Pro Leu Leu Ser Ser Leu Ser
100 105 110
Pro Pro Leu Ser Ala Ile Val Ser Asp Phe Thr Leu Thr Ser Gln Val
115 120 125
Thr Asp Leu Val Ser Asp Leu Pro Ile Ser Thr Tyr Thr Leu Met Thr
130 135 140
Ser Ser Ala Ala Phe Phe Cys Leu Met Ala Tyr Leu Pro Lys Leu Leu
145 150 155 160
Gln Ile Asp Val Ala Asn Arg Asp Ala Ile Glu Ile Pro Asp Leu Gly
165 170 175
Pro Ile Ser Met Ser Ser Ile Pro Pro Lys Met Leu Asp Pro Ser Asp
180 185 190
Phe Phe Ser Ala Phe Ile Ser Ser Asn Val Ser Ser Leu His Lys Val
195 200 205
Lys Gly Val Leu Ile Asn Thr Phe Asn Ser Phe Glu Ser Glu Ala Ile
210 215 220
Glu Ala Val Arg Arg Asn Gly Val Asp His Ile Leu Pro Ile Gly Pro
225 230 235 240
Leu Glu Ser Tyr Asp Ala Lys Lys Ala His Asp Leu Pro Trp Leu Asp
245 250 255
Glu Gln Pro Pro Glu Ser Val Leu Phe Val Ser Phe Gly Ser Arg Thr
260 265 270
Ala Leu Ser Lys Glu Gln Ile Arg Glu Leu Gly Ala Ala Leu Glu Lys
275 280 285
Ser Gly Cys Arg Phe Leu Trp Val Leu Lys Gly Gly Lys Val Asp Lys
290 295 300
Glu Asp Lys Glu Glu Val Glu Asp Met Leu Gly Ala Ser Phe Val Glu
305 310 315 320
Arg Thr Lys Lys Lys Gly Leu Ile Val Lys Gly Trp Val Lys Gln Glu
325 330 335
Gln Ile Leu Ala His Pro Ala Ile Gly Gly Phe Val Ser His Cys Gly
340 345 350
Trp Asn Ser Val Ile Glu Ala Ala Arg Leu Gly Val Pro Val Leu Ala
355 360 365
Trp Pro Gln His Gly Asp Gln Ser Val Asn Ala Gly Val Val Glu Lys
370 375 380
Ala Gly Leu Gly Leu Trp Val Arg Glu Trp Gly Trp Gly Gln Thr Lys
385 390 395 400
Leu Ile Gly Arg Glu Glu Ile Ala Glu Lys Met Ile Glu Val Met Gln
405 410 415
Asp Glu Lys Leu Arg Val Ser Ala Gly Glu Val Arg Ala Lys Ala Lys
420 425 430
Glu Thr Arg Glu Val Asp Gly Asp Ser Glu Ala Leu Leu Gln Arg Leu
435 440 445
Ile His Ser Phe Asn Asn Ile Thr Gln Asn Ser
450 455
<210> 15
<211> 458
<212> PRT
<213> 黄芩 (Scutellaria baicalensis)
<400> 15
Met Ser Lys Ser Glu Asn Ala Gly Gln Arg Pro His Val Ala Val Phe
1 5 10 15
Pro Cys Ala Gly Met Gly His Leu Leu Pro Tyr Leu Arg Leu Ala Ala
20 25 30
Met Leu His Ser Arg Gly Cys Ala Val Ser Val Ile Ser Ala His Pro
35 40 45
Thr Ile Ser Asp Ala Glu Ser Arg Ser Leu Ser Ser Phe Phe Ser Leu
50 55 60
Tyr Pro Gln Ile Arg Ser Leu Glu Ile Gln Leu Leu Pro Leu Lys Arg
65 70 75 80
Asn Pro Arg Phe Thr Asn Asp Asp Pro Phe Phe Ile Gln Arg Glu Ser
85 90 95
Ile Gly Asn Ser Ile His Leu Leu Arg Pro Leu Leu Ala Ser Leu Ser
100 105 110
Pro Pro Leu Ser Ala Ile Phe Val Asp Phe Pro Val Leu Thr Glu Phe
115 120 125
Ser Pro Ile Ala Ala Asp Phe Ser Leu Pro Thr Tyr Thr Leu Ile Val
130 135 140
Thr Ser Ala Arg Phe Phe Ser Leu Met Ala His Leu Pro Arg Leu Leu
145 150 155 160
Glu Gln Glu Asp Asp Ile Ser Lys Lys Ser Glu Val Cys Val Pro His
165 170 175
Leu Asp Pro Ile Gln Val Ser Ser Ile Pro Pro Gln Met Leu Asp Arg
180 185 190
Arg His Phe Phe Val Glu Thr Ile Thr Ser Asn Val Ala Ser Leu Ser
195 200 205
Tyr Leu Lys Gly Val Leu Ile Asn Thr Phe Thr Trp Leu Glu Pro Glu
210 215 220
Ala Val Glu Ala Leu Lys Arg Asn Gly Val Asp His Ile Leu Pro Ile
225 230 235 240
Gly Pro Leu Glu Ala Ile Lys Ala Glu Glu Ser Asp Met Asp Leu Pro
245 250 255
Trp Leu Glu Glu Gln Ala Pro Lys Ser Val Leu Phe Ile Ser Phe Gly
260 265 270
Ser Arg Gly Ala His Thr Lys Glu Gln Leu Arg Glu Phe Ala Ala Ala
275 280 285
Leu Glu Lys Ser Gly Trp Arg Phe Leu Trp Val Leu Lys Ser Gly Lys
290 295 300
Val Asp Arg Glu Asp Lys Glu Glu Thr Glu Asp Ile Leu Gly Ser Ser
305 310 315 320
Phe Leu Glu Arg Thr Lys Asn Arg Gly Val Val Ile Lys Gly Trp Ala
325 330 335
Asp Gln Glu Arg Ile Leu Ala His Ser Ala Ile Gly Gly Phe Val Ser
340 345 350
His Cys Gly Trp Asn Ser Val Val Glu Ala Ala Lys Leu Gly Val Pro
355 360 365
Val Leu Ala Trp Pro Pro His Gly Asp Gln Arg Val Asn Ala Glu Val
370 375 380
Val Glu Lys Val Glu Leu Gly Leu Trp Val Arg Gly Trp Gly Trp Ala
385 390 395 400
Gly Glu Arg Leu Ile Gly Arg Asp Glu Ile Ala Glu Lys Leu Ile Glu
405 410 415
Leu Arg Asn Asp Glu Arg Leu Arg Glu Arg Val Lys Glu Val Arg Glu
420 425 430
Lys Ala Arg Glu Glu Arg Glu Ser Gly Gly Ile Ser Glu Thr Leu Ile
435 440 445
Arg Asp Leu Ile His Ser Leu Lys Ile Lys
450 455
<210> 16
<211> 456
<212> PRT
<213> 水稻 (Oryza sativa)
<400> 16
Met Ala Pro Pro Thr Val Leu Asn Ser Gly Glu Pro Asp Ala Arg Arg
1 5 10 15
Arg Glu Arg Pro His Val Val Phe Ile Pro Ser Ala Gly Met Gly His
20 25 30
Leu Leu Pro Phe Phe Arg Phe Ile Gly Ala Leu Ser Ser His Asp Val
35 40 45
Asp Ile Ser Val Val Thr Val Leu Pro Thr Val Ser Ala Ala Glu Ala
50 55 60
Asp His Phe Ala Arg Leu Phe His Asp Phe Pro Ser Ile Arg Arg Val
65 70 75 80
Asp Phe Asn Leu Leu Pro Leu Asp Ala Ser Glu Phe Pro Gly Ala Asp
85 90 95
Pro Phe Leu Leu Arg Trp Glu Ala Leu Arg Arg Ser Met His Leu Leu
100 105 110
Ala Pro Ala Ile Ala Gly Val Ala Pro Arg Ala Thr Ala Val Val Thr
115 120 125
Asp Val Thr Leu Val Ser His Val Asn Pro Ile Ala Lys Asp Leu Arg
130 135 140
Leu Gln Cys His Asp Ala Gln Ala Asp Val Gly Asp Val Asp Val Pro
145 150 155 160
Gly Val Arg His Leu Gln Arg Ser Trp Leu Pro Gln Pro Leu Leu Asp
165 170 175
Leu Asp Met Leu Phe Thr Lys Gln Phe Ile Glu Asn Gly Arg Glu Val
180 185 190
Val Lys Thr Asp Gly Val Leu Ile Asn Thr Phe Asp Ala Leu Glu Pro
195 200 205
Val Ala Leu Ala Ala Leu Arg Asp Gly Thr Val Val Arg Gly Phe Pro
210 215 220
Pro Val Phe Ala Val Gly Pro Tyr Ser Ser Leu Ala Ser Glu Lys Lys
225 230 235 240
Ala Ala Asp Ala Asp Gln Ser Ser Ala Leu Ala Trp Leu Asp Gln Gln
245 250 255
Pro Ala Arg Ser Val Val Tyr Val Ala Phe Gly Asn Arg Cys Thr Val
260 265 270
Ser Asn Asp Gln Leu Arg Glu Ile Ala Ala Gly Leu Glu Ala Ser Gly
275 280 285
Cys Arg Phe Leu Trp Ile Leu Lys Thr Thr Val Val Asp Arg Asp Glu
290 295 300
Ala Ala Ala Gly Gly Val Arg Asp Val Leu Gly Asp Gly Phe Met Glu
305 310 315 320
Arg Val Lys Gly Arg Gly Met Val Thr Lys Glu Trp Val Asp Gln Glu
325 330 335
Ala Val Leu Gly His Pro Ala Val Gly Leu Phe Leu Ser His Ser Gly
340 345 350
Trp Asn Ser Val Thr Glu Ala Ala Ala Ala Gly Val Pro Leu Leu Ala
355 360 365
Trp Pro Arg Gly Gly Asp His Arg Val Ala Ala Thr Val Val Ala Ser
370 375 380
Ser Gly Val Gly Val Trp Met Glu Gln Trp Ser Trp Asp Gly Glu Glu
385 390 395 400
Trp Leu Val Ser Gly Glu Glu Ile Gly Gly Lys Val Lys Glu Met Met
405 410 415
Ala Asp Asp Ala Val Arg Glu Arg Ala Ala Lys Val Gly Glu Glu Ala
420 425 430
Ala Lys Ala Val Ala Glu Gly Gly Thr Ser His Thr Ser Met Leu Glu
435 440 445
Phe Val Ala Lys Leu Lys Ala Ala
450 455
<210> 17
<211> 481
<212> PRT
<213> 水稻 (Oryza sativa)
<400> 17
Met Ala Pro Pro Pro Ala Val Leu Ser Ser Gly Glu Leu Gly Ala Arg
1 5 10 15
Gly His Thr Arg Pro His Val Val Phe Val Pro Ser Ala Gly Met Gly
20 25 30
His Leu Leu Gln Phe Phe Arg Phe Ile Gly Ala Leu Ser Ala His Asp
35 40 45
Val Asp Ile Ser Val Val Thr Val Phe Pro Thr Val Ser Ala Ala Glu
50 55 60
Ala Asp His Phe Ala Ala Leu Phe Arg Asp Tyr Pro Ser Val Arg Arg
65 70 75 80
Leu Asp Phe Asp Leu Leu Pro Phe Asp Ala Ser Glu Phe Pro Gly Gly
85 90 95
Asp Pro Phe Leu Leu Arg Trp Glu Ala Leu Arg Arg Ser Leu His Leu
100 105 110
Leu Gly Pro Val Ile Ala Gly Val Thr Pro Arg Val Thr Ala Thr Val
115 120 125
Thr Asp Val Thr Leu Val Ser His Val Asn Pro Ile Ala Lys Asp Leu
130 135 140
Gly Val Gln Cys His Val Leu Tyr Val Ser Ser Ala Ala Met Met Ser
145 150 155 160
Leu Cys Ser Tyr Phe Pro Ile Tyr Leu Asp Asn Lys Asp Ala Gly Ala
165 170 175
Asp Val Gly Asp Val Asp Ile Pro Gly Val Arg Arg Leu Lys Arg Ser
180 185 190
Trp Leu Pro Gln Pro Leu Leu Asp Leu Asn Lys Leu Phe Thr Lys Gln
195 200 205
Phe Ile Glu Asn Gly Arg Glu Met Val Lys Thr Asp Gly Val Leu Ile
210 215 220
Asn Thr Phe Asp Ala Leu Glu Pro Val Ala Leu Ala Ala Leu Arg Asp
225 230 235 240
Gly Lys Val Val Arg Gly Phe Pro Pro Val Phe Ala Val Gly Pro His
245 250 255
Ser Ser Leu Ala Ser Glu Ala Thr Lys Gly Ala Ala Ala Asp Ala Glu
260 265 270
Gly Ser Pro Met Ala Trp Leu Arg Gln Gln Pro Ala Arg Ser Val Val
275 280 285
Tyr Val Ala Phe Gly Ser Arg Cys Ala Val Ser His Glu Gln Ile Arg
290 295 300
Glu Ile Ala Ala Gly Leu Glu Ala Ser Gly Ser Arg Phe Leu Trp Ile
305 310 315 320
Leu Lys Thr Thr Val Val Asp Arg Asp Asp Asp Ala Gly Ile Arg Asp
325 330 335
Val Leu Gly Asp Gly Phe Leu Glu Arg Val Arg Gly Arg Gly Val Val
340 345 350
Thr Lys Ala Trp Val Asp Gln Asp Ala Val Leu Arg Asp Pro Ala Val
355 360 365
Gly Leu Phe Leu Ser His Ser Gly Trp Asn Ser Val Ile Glu Ala Ala
370 375 380
Thr Ala Gly Val Pro Leu Leu Ala Trp Pro Arg Gly Gly Asp His Arg
385 390 395 400
Val Ala Ala Thr Val Val Ala Ser Ser Gly Val Gly Val Trp Met Glu
405 410 415
Gln Trp Ser Trp Asp Gly Glu Glu Trp Val Val Ser Gly Glu Glu Ile
420 425 430
Gly Gly Lys Val Lys Glu Met Met Ala Asp Ala Gly Val Arg Glu Lys
435 440 445
Ala Ala Lys Val Gly Glu Glu Val Ala Lys Ala Val Ala Val Gly Gly
450 455 460
Thr Ser His Thr Gly Ile Leu Asp Phe Val Ala Lys Leu Lys Ala Ala
465 470 475 480
Thr
<210> 18
<211> 472
<212> PRT
<213> 甘草 (Glycyrrhiza uralensis）
<400> 18
Met Gly Glu Asn Asn Met Ser Ser Ile Ala Pro His Pro Val Val His
1 5 10 15
Val Ala Leu Leu Pro Ser Ala Gly Met Gly His Leu Ser Pro Phe Leu
20 25 30
Arg Leu Ala Ser Leu Leu Leu His Gln His Cys His Val Thr Leu Ile
35 40 45
Thr Pro Gln Leu Thr Val Ser Lys Ala Glu Glu Asp Leu Leu Ser Arg
50 55 60
Phe Leu Ser Ala Phe Pro Gln Val Asn Gln Leu His Phe His Leu Pro
65 70 75 80
Pro Ser Asp Ser Thr Ile Ser Thr Asp Pro Phe Phe Leu Gln Phe Ala
85 90 95
Ser Ile Arg Ser Ser Ser His Leu Leu Thr Pro Leu Leu Ser Ser Leu
100 105 110
Thr Pro Pro Leu Ser Ser Phe Ile Tyr Asp Met Thr Leu Ile Ser Pro
115 120 125
Leu Leu Pro Ile Ala Glu Ser Leu Gly Val Pro His Tyr Ile Leu Phe
130 135 140
Thr Ser Ser Ala Thr Met Phe Ser Phe Phe Ser Tyr Phe Pro Thr Leu
145 150 155 160
Ala Lys Ser Glu Ser Phe Pro Gly Lys Leu Asp Phe Val Glu Ile Pro
165 170 175
Gly Val Ser Val Ser Ser Ile Pro Arg Ser Ser Ile Pro Pro Pro Leu
180 185 190
Leu Val Pro Asn Ser Leu Phe Gly Lys Leu Phe Met Glu Asp Ser Pro
195 200 205
Lys Leu Lys Lys Leu His Gly Val Leu Val Asn Thr Phe Glu Gly Ile
210 215 220
Glu Lys Leu Ser Leu Glu Ala Leu Asn Gly Gly Lys Val Val Lys Gly
225 230 235 240
Leu Pro Pro Val Tyr Gly Val Gly Pro Phe Val Pro Cys Glu Phe Glu
245 250 255
Lys Val Val Lys Arg Gly Glu Thr Ile Ser Glu Trp Leu Asp Glu Gln
260 265 270
Pro Ser Gly Ser Val Val Tyr Val Ser Phe Gly Ser Arg Thr Ala Met
275 280 285
Gly Arg Glu Gln Leu Arg Glu Val Gly Asp Gly Leu Val Lys Ser Gly
290 295 300
Trp Arg Phe Leu Trp Val Val Lys Asp Lys Ile Val Asp Arg Ala Glu
305 310 315 320
Glu Glu Gly Leu Asp Gly Val Leu Gly Phe Glu Leu Val Glu Arg Met
325 330 335
Val Lys Glu Lys Lys Gly Leu Val Val Lys Glu Trp Val Asp Gln Ser
340 345 350
Glu Ile Leu Gly His Lys Ala Val Gly Gly Phe Val Ser His Cys Gly
355 360 365
Trp Asn Ser Val Val Glu Ala Ala Trp Phe Gly Val Lys Ile Leu Gly
370 375 380
Trp Pro Leu His Gly Asp Gln Lys Ile Asn Ala Glu Val Val Ala Lys
385 390 395 400
Gly Gly Trp Gly Val Trp Lys Glu Gly Trp Asp Trp Glu Gly Glu Arg
405 410 415
Leu Val Lys Gly Glu Glu Ile Gly Glu Ala Ile Arg Glu Val Met Asn
420 425 430
Asp Glu Ser Leu Val Met Lys Ala Thr Gln Val Lys Lys Asp Ala Arg
435 440 445
Lys Ala Ile Ser Val Gly Gly Gly Cys Glu Val Ala Leu Gln Lys Leu
450 455 460
Met Glu Val Trp Lys Lys Asn Val
465 470
<210> 19
<211> 485
<212> PRT
<213> 甘草 (Glycyrrhiza uralensis）
<400> 19
Met Thr Ser Cys Thr Asn Ser Glu Pro Val Thr Val His Val Ala Phe
1 5 10 15
Leu Pro Ser Ala Gly Met Gly His Leu Asn Pro Cys Leu Arg Thr Ala
20 25 30
Glu Gln Phe Leu Arg His Gly Cys Lys Val Thr Leu Ile Thr Pro Lys
35 40 45
Pro Thr Val Ser Leu Ala Glu Ser Asn Leu Ile Asn Arg Phe Cys Ser
50 55 60
Ser Phe Pro Ser Ser Gln Leu Asn Gln Val Asp Leu Asn Leu Ile Asp
65 70 75 80
Val Asp Pro Thr Gln Val Asn Thr Asn Asp Pro Phe Trp Leu His Phe
85 90 95
Glu Thr Ile Arg Gln Ser Leu Gln Tyr His Leu Pro Thr Ile Leu Ser
100 105 110
Ser Leu Ser Ser Pro Pro Leu Ser Ala Phe Ile Tyr Asp Val Ser Leu
115 120 125
Ile Ser Pro Leu Val Pro Ile Thr Glu Lys Leu Cys Phe Pro Ser Tyr
130 135 140
Ile Tyr Phe Ile Ala Pro Ala Arg Met Phe Ser Phe Phe Ala His Leu
145 150 155 160
Ser Val Leu Ser Glu Lys Asn Pro Gly Val Lys Pro Ser Ser Phe Ile
165 170 175
Gly Gly Val Val Glu Ile Pro Gly Ile Ala Pro Ile Pro Arg Ser Ser
180 185 190
Val Pro Pro Leu Leu Leu Glu Pro Asn Ser Leu Phe Glu Ser Ile Phe
195 200 205
Met Glu Asp Ser Pro Lys Leu Lys Lys Leu Ser Gly Ile Phe Ile Asn
210 215 220
Ser Phe Glu Glu Leu Glu Pro Glu Ala Leu Ala Ala Leu Asn Asn Gly
225 230 235 240
Arg Val Val Pro Gly Leu Pro Pro Val Tyr Pro Ile Gly Pro Leu Val
245 250 255
Pro Cys Glu Phe Glu Lys Val Arg Ser Ser Pro Lys Thr Asn Ser Ser
260 265 270
Thr Ile Leu Lys Trp Leu Asp Gly Gln Pro Ala Gly Ser Val Val Tyr
275 280 285
Val Cys Leu Gly Asn Arg Thr Gln Thr Arg Arg Glu Gln Ile Lys Asp
290 295 300
Met Ala Gln Gly Leu Ile Ser Cys Gly Tyr Lys Phe Leu Trp Val Val
305 310 315 320
Lys Leu Lys Ile Val Asp Arg Glu Asp Gln Glu Glu Glu Leu Asp Asp
325 330 335
Val Leu Gly Asn Glu Val Met Lys Gln Val Asn Glu Lys Gly Met Ala
340 345 350
Val Lys Thr Trp Val Asp Gln Met Glu Ile Leu Gly His Pro Ser Val
355 360 365
Gly Gly Phe Val Asn His Gly Gly Trp Asn Ser Leu Val Glu Gly Val
370 375 380
Trp His Gly Lys Pro Ile Leu Ser Trp Ala His Asp Gly Asp Gln Lys
385 390 395 400
Ile Ala Ser Glu Val Val Lys Met Asn Gly Val Gly Ile Trp Pro Gln
405 410 415
Val Trp Gly Trp Gly Ala Arg Asp Asp Val Val Lys Gly Glu Glu Ile
420 425 430
Ala Arg Ala Ile Asn Glu Met Met Ser Asn Glu Ser Leu Arg Asn Lys
435 440 445
Ala Arg Glu Met Lys Glu Ala Ala Arg Lys Ala Ala Gly Val Gly Gly
450 455 460
Ser Cys Glu Val Ile Ile Lys Arg Gln Ile Glu Glu Trp Lys Lys Arg
465 470 475 480
Ile Val Gln Gly Ile
485
<210> 20
<211> 473
<212> PRT
<213> 一把伞南星 (Arisaema erubescens)
<400> 20
Met Ser Pro Pro Thr Ala Glu Gly Ala Arg Thr Pro Arg Val Ala Leu
1 5 10 15
Leu Pro Ser Ala Gly Met Gly His Leu Thr Pro Phe Leu Arg Leu Ala
20 25 30
Ala Val Leu Ala Ala Arg Gly Cys Asp Val Thr Leu Ile Thr Pro His
35 40 45
Pro Val Val Ser Ala Ala Glu Ala Arg His Ile Ala Ala Phe Leu Ala
50 55 60
Ser Ala Ser Pro Arg Val Arg Ser Leu Glu Phe Arg Leu Leu Pro Phe
65 70 75 80
Asp Ala Ala Ser Ala Thr Ser Thr Asp Pro Phe Phe Leu Gln Phe Glu
85 90 95
Ala Ile Arg Arg Ser Ala His Leu Leu Gly Pro Leu Leu Ala Ser Ala
100 105 110
Cys Pro Ala Leu Ser Ala Val Val Thr Asp Val Ser Leu Ile Ser Ala
115 120 125
Val Ala Pro Val Val Ala Ala Val Gly Leu Pro Asn Tyr Val Ile Phe
130 135 140
Thr Ser Ser Ala Ala Met Leu Ala Leu Cys Val Ala Phe Pro Ser Ala
145 150 155 160
Ser Ala Ser Leu Gly Gly Ala Ala Asp Thr Phe Glu Val Pro Gly Leu
165 170 175
Ala Ala Leu Pro Ile Asn Trp Leu Pro Gln Pro Leu Arg Asp Pro Gly
180 185 190
Asn Leu Phe Thr Thr Gln Phe Val Glu Asn Gly Arg Glu Leu Thr Arg
195 200 205
Val Asp Gly Ile Met Ile Asn Thr Trp Asp Ala Leu Glu Gly Glu Thr
210 215 220
Leu Ala Ala Leu Asn Ala Gly Ala Val Ala His Leu Pro Pro Val Ile
225 230 235 240
Pro Val Gly Pro Leu Pro Pro Val Lys Pro Arg Ser Ile Pro Gly Ile
245 250 255
Pro Trp Leu Asp Asp Gln Pro Glu Arg Ser Val Val Tyr Val Ser Phe
260 265 270
Gly Ser Arg Thr Ala Leu Ser Ala Glu Gln Ile Arg Glu Leu Gly Ala
275 280 285
Gly Leu Glu Lys Ser Gly Cys Arg Phe Leu Trp Ala Ala Lys Ser Lys
290 295 300
Lys Val Asp Lys Glu Asp Asp Gly Val Val Leu Glu Glu Leu Leu Gly
305 310 315 320
Asp Asp Gly Phe Leu Arg Arg Met Glu Gly Arg Gly Leu Val Val Lys
325 330 335
Asp Trp Val Asp Gln Asp Ala Val Leu Ala His Pro Ala Val Gly Gly
340 345 350
Phe Leu Cys His Cys Gly Trp Asn Ser Val Thr Glu Ala Ala Ile His
355 360 365
Gly Val Arg Val Leu Ala Trp Pro Arg His Gly Asp Gln Arg Leu Asn
370 375 380
Ala Trp Val Val Glu Arg Gly Gly Leu Gly Thr Trp Pro Thr Glu Trp
385 390 395 400
Ser Trp Glu Gly Asp Ala Glu Leu Val Ser Gly Glu Glu Ile Ala Arg
405 410 415
Arg Val Arg Glu Leu Val Gly Ser Pro Asp Ala Ser Asp Ala Ala Ala
420 425 430
Pro Val Gly Glu Glu Ala Val Arg Ala Ala Glu Ala Cys Gly Ser Ser
435 440 445
Arg Lys Ala Leu Asp Glu Leu Ile Glu Lys Trp Gly Arg Gln Leu Gly
450 455 460
Thr Arg Ser Asp Gly Met Arg Arg Pro
465 470
<210> 21
<211> 485
<212> PRT
<213> 一把伞南星 (Arisaema erubescens)
<400> 21
Met Ser Arg Glu Gln Gln Ser Ser Ser Ser Asn Gly Gly Leu Thr Val
1 5 10 15
Pro His Val Ala Val Ile Pro Ser Ala Gly Met Gly His Leu Ser Ile
20 25 30
Ser Val Arg Leu Ala Ala Thr Leu Ala Ser His Gly Cys Leu Val Thr
35 40 45
Phe Ile Thr Pro Arg Pro Val Val Ser Asp Ala Glu Ser Arg Thr Ile
50 55 60
Ala Asp Leu His Ser Ser Tyr Pro Gln Ile Arg Ser Leu Asp Leu Arg
65 70 75 80
Leu Leu Pro Phe Asp Ala Pro Asp Val Asp Pro Phe Leu Arg Gln Phe
85 90 95
Glu Val Ile Arg Arg Ser Ala Ser Leu Leu Gln Pro Leu Leu Thr Ser
100 105 110
Val Thr Pro Pro Leu Ser Ala Val Ile Val Asp Ile Ser Leu Leu Ser
115 120 125
Thr Phe Val Pro Val Thr Lys Ala Leu Arg Leu Pro Thr Tyr Val Thr
130 135 140
Phe Ile Ser Cys Ala Met Met Leu Ala Leu Leu Ala Val Phe Pro Ser
145 150 155 160
Val Arg Lys Ala Ala Gly Asp Gly Pro Ile Ala Ala Val Asp Ile Pro
165 170 175
Gly Val Gly Thr Ser Leu Pro Gly Ser Trp Ile Pro Pro Pro Leu His
180 185 190
Asp Pro Lys His Leu Phe Ala Ile Gln Phe Leu Glu Asn Gly Glu Ala
195 200 205
Leu Leu Gln Ala Asp Gly Val Phe Ile Asn Thr Phe Ser Ala Leu Glu
210 215 220
Ala Glu Thr Leu Ala Ala Leu Asn Ala Gly Asn Val Val Pro Gly Leu
225 230 235 240
Pro Pro Val Ile Ala Ile Gly Pro Gln Gln Pro Val Lys Phe Glu Gly
245 250 255
Ala Lys Arg Ala Ala Ala Ala Gly Arg Asp Glu Trp Ala Ala Trp Leu
260 265 270
Asp Ala Gln Pro Pro Gly Ser Val Leu Tyr Val Ala Phe Gly Asn Arg
275 280 285
Thr Ala Leu Pro Arg Glu Gln Leu Arg Glu Leu Gly Leu Gly Leu Glu
290 295 300
Lys Ser Gly Cys Arg Phe Leu Trp Val Val Lys Ser Lys Ile Val Asp
305 310 315 320
Arg Glu Asp Asp Ala Gly Val Leu Leu Glu Asp Leu Leu Gly Glu Gly
325 330 335
Tyr Arg Glu Arg Val Gly Glu Arg Gly Met Val Val Lys Ser Trp Val
340 345 350
Asp Gln Glu Glu Val Leu Ala His Pro Ser Val Gly Gly Phe Leu Ser
355 360 365
His Ala Gly Trp Asn Ser Val Ala Glu Ala Ala Leu Gln Gly Ile Cys
370 375 380
Leu Phe Gly Trp Pro Ala Gly Gly Asp Gln Arg Val Ala Ala Leu Val
385 390 395 400
Val Glu Lys Cys Gly Ala Gly Ala Trp Val Lys Glu Trp Gly Trp Ser
405 410 415
Gly Ser Gly Arg Gly Leu Glu Glu Thr Pro Thr Val Ser Gly Glu Glu
420 425 430
Ile Gly Lys Arg Leu Arg Glu Leu Met Gly Asp Ala Ser Leu Arg Glu
435 440 445
Ser Thr Ala Arg Ile Arg Lys Asp Ala Leu Arg Ser Val Gly Val Gly
450 455 460
Gly Lys Ser Tyr Glu Ala Val Arg Ser Phe Ile Ala Gly Leu Gly Gly
465 470 475 480
Val Gly Tyr Thr Thr
485
<210> 22
<211> 484
<212> PRT
<213> 少根紫萍 (Landoltia punctate)
<400> 22
Met Arg Pro Gly Ser Ser Glu Arg Asn His Pro Ala Glu Arg Pro Thr
1 5 10 15
Thr Ile Leu Ser Gly Ser Met Ala Pro Ala Ala Val Pro His Val Ala
20 25 30
Leu Leu Pro Ser Ser Gly Met Gly His Leu Thr Pro Phe Leu Arg Leu
35 40 45
Ala Ala Ala Leu Ala Ser His Gly Cys Val Ile Thr Phe Ile Thr Pro
50 55 60
Thr Pro Val Val Ser Ala Ala Glu Ala Arg His Val Glu Ala Phe Ile
65 70 75 80
Ser Ser Ser Pro Leu Phe Arg Arg Leu Glu Phe Pro Leu Leu Pro Phe
85 90 95
Asp Val Ser Ser Val Val Ser Asp Asp Pro Phe Phe Leu Gln Phe Glu
100 105 110
Arg Ile Ser Arg Ser Ala Arg His Leu Gly Pro Val Leu Ser Ser Val
115 120 125
Ser Pro Pro Leu Ser Ala Leu Ile Leu Asp Val Thr Leu Thr Ser Ser
130 135 140
Ile Leu Pro Ile Ala Ala Glu Ile Ser Leu Pro Ala Tyr Ile Leu Phe
145 150 155 160
Thr Ser Ser Ala Gly Met Leu Ser Leu Cys Leu Ser Tyr Pro Glu Ile
165 170 175
Ala Ala Ser Gly Gly Val Thr Ile Lys Ile Pro Gly Val Ala Glu Asp
180 185 190
Leu Ala Pro Ser Ser Leu Pro Gln Pro Leu Arg Asp Pro Arg Asn Leu
195 200 205
Phe Thr Gly Gln Phe Ile Glu Asn Gly Arg Ala Met Ala Arg Ala Asp
210 215 220
Gly Ile Ile Ile Asn Thr Trp Glu Ala Leu Glu Pro Ala Thr Leu Ala
225 230 235 240
Ala Leu Gln Gly Ser Lys Ala Val Ser Gly Phe Pro Leu Val Ile Pro
245 250 255
Val Gly Pro Leu Leu Ala Ala Ser Asp Ile His Thr Ala Ala Asp Asp
260 265 270
Leu Val Ile Pro Trp Leu Asp Ala Gln Pro Ala Ser Ser Val Val Phe
275 280 285
Val Ser Phe Gly Ser Arg Thr Ala Leu Ser Ala Glu Gln Leu Arg Glu
290 295 300
Leu Ala Ala Gly Leu Glu Ser Ser Gly Cys Arg Phe Leu Trp Val Leu
305 310 315 320
Lys Thr Lys Lys Val Asp Arg Glu Asp Gln Glu Gly Glu Lys Ala Val
325 330 335
Asp Glu Leu Leu Gly Glu Gly Phe Leu Gln Arg Val Glu Trp Lys Gly
340 345 350
Lys Val Val Ser Gly Trp Val Asp Gln Arg Ala Val Leu Asp His Pro
355 360 365
Ser Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Val Thr Glu
370 375 380
Ala Ala Leu Gly Gly Met Arg Val Leu Ala Trp Pro Arg His Gly Asp
385 390 395 400
Gln Arg Ile Asn Ala Met Val Val Glu Lys Ser Gly Leu Gly Lys Trp
405 410 415
Pro Ser Leu Trp Thr Trp Glu Gly Asp Asp Glu Ile Val Arg Arg Glu
420 425 430
Glu Ile Ala Gly Arg Val Ala Glu Leu Met Ala Ser Pro Ala Ala Ala
435 440 445
Ala Ala Ala Ala Lys Val Lys Glu Glu Ala Val Arg Ala Ala Thr Ala
450 455 460
Gly Gly Ser Ser Gln Arg Gln Leu Glu Asp Leu Val Ser Arg Phe Thr
465 470 475 480
Cys Ser Gly Asp
<210> 23
<211> 468
<212> PRT
<213> 少根紫萍 (Landoltia punctate)
<400> 23
Met Ser Pro Pro Ala Pro Ala Asp Val Val Ser Ser Ala Lys Pro His
1 5 10 15
Val Ala Val Ile Pro Ala Ala Gly Met Gly His Leu Asn Pro Thr Leu
20 25 30
Arg Leu Ala Gly Glu Leu Ala Ser Arg Gly Cys Val Val Thr Phe Ile
35 40 45
Asn Pro Ser Pro Pro Val Ser Leu Ala Glu Ala Thr Ser Val Ala Glu
50 55 60
Phe Val Ala Ser Thr Pro Gly Val Arg Leu Leu Asp Leu Pro Val Gln
65 70 75 80
Pro Leu Asp Pro Ser Cys Phe Pro Ala His Glu Asp Pro Phe Leu Arg
85 90 95
Gln Phe Glu Ala Val Arg Arg Ser Ala Pro Leu Leu Thr Pro Leu Leu
100 105 110
Ser Asp Val Ser Pro Pro Leu Ala Ala Ile Val Cys Asp Ile Ala Ile
115 120 125
Cys Ser Thr Phe Leu Thr Val Ala Ala Glu Ile Ser Leu Pro Ala Tyr
130 135 140
Val Phe Phe Ser Leu Ser Ala Gln Met Leu Ser Leu Asn Leu Ala Phe
145 150 155 160
Pro Thr Val Ala Asp Gln Val Tyr Gly Ala Gly Glu Gly Asp Glu Ile
165 170 175
Arg Phe Pro Gly Leu Pro Glu Ser Ile Pro Arg Ser Trp Leu Pro Pro
180 185 190
Pro Leu Leu Asp Pro Ala His Leu Phe Ala Val His Phe Val Glu Asn
195 200 205
Gly Lys Ala Met Pro Arg Ala Ala Gly Ile Leu Val His Ser Trp Glu
210 215 220
Ala Leu Glu Pro Glu Ala Leu Ala Ala Leu Arg Gly Gly Arg Val Leu
225 230 235 240
Ala Gly Leu Pro Pro Val Leu Pro Ile Gly Pro Leu Tyr Gln Lys Glu
245 250 255
Lys Ser Asn Ala Val Phe Leu Pro Trp Leu Asp Ala Gln Arg Asp Arg
260 265 270
Ser Val Leu Phe Val Cys Phe Gly Asn Arg Ser Thr His Ser Pro Glu
275 280 285
Gln Leu Arg Glu Met Ala Ala Gly Leu Glu Arg Ser Gly Cys Arg Phe
290 295 300
Val Trp Val Leu Lys Thr Lys Val Val Asp Lys Asp Glu Asp Glu Gly
305 310 315 320
Ala Gln Lys Glu Ile Leu Gly Glu Gly Tyr Leu Glu Arg Val Lys Glu
325 330 335
Arg Gly Val Val Ile Asn Gly Trp Val Asp Gln Met Thr Ile Leu Ser
340 345 350
His Arg Ala Val Gly Gly Phe Phe Ser His Ser Gly Ser Ser Ser Val
355 360 365
Ala Glu Ala Ala Ile Gly Gly Gln Pro Leu Leu Leu Trp Pro Met Gly
370 375 380
Gly Asp Gln Arg Met Ser Ala Leu Val Ala Glu Arg Arg Gly Met Gly
385 390 395 400
Val Trp Pro Arg Gly Trp Gly Trp Ser Ala Asp Asp Lys Leu Ile Pro
405 410 415
Gly Glu Glu Ile Ala Arg Arg Ile Lys Asp Phe Met Gly Asp Asn Ala
420 425 430
Leu Arg Ala Val Ala Ala Lys Met Lys Lys Glu Thr Ala Ser Ala Met
435 440 445
Ala Pro Gly Gly Ser Lys Asp Gln Trp Phe Asp Asp Phe Ile Ala Arg
450 455 460
Ile Asn Arg Val
465
<210> 24
<211> 494
<212> PRT
<213> 大薸 (Pistia stratiotes)
<400> 24
Met Val Gln Glu Thr Ser Pro Pro Val Cys Asn Phe Arg Asp Thr Val
1 5 10 15
Ser Thr Thr Ser Pro Ala Pro His Ile Ala Ile Leu Ser Ser Ala Gly
20 25 30
Phe Gly His Leu Ile Val Ser Val Arg Leu Ala Val Ala Leu Ala Ser
35 40 45
His Gly Cys Leu Ile Thr Leu Val Thr Pro Leu Pro Thr Val Ser Glu
50 55 60
Ala Glu Ser Arg Tyr Ile Ser Asp Phe His Ser Cys Phe Pro His Ile
65 70 75 80
Arg Leu Leu Glu Phe His Leu Leu Pro Leu Asp Ala Gln Ala Asp Thr
85 90 95
Asp Pro Phe Phe Ser Gln Phe Glu Thr Ile Arg Arg Ser Ala Ala Leu
100 105 110
Leu Gly Pro Leu Leu Ala Ser Ala Thr Pro Pro Leu Ser Ala Leu Leu
115 120 125
Ala Asp Ile Ser Val Leu Ser Ser Tyr Ile Pro Val Thr Lys Ala Leu
130 135 140
Gly Ile Pro Thr Tyr Val Thr Phe Thr Ser Cys Ala Ala Met Leu Ala
145 150 155 160
Leu Ser Cys Val Phe Pro Arg Val Arg Lys Glu Val Gly Asp Gly Pro
165 170 175
Val Val Ser Ile Asp Ile Pro Gly Leu Gly Thr Val Pro Gly Ser Trp
180 185 190
Ile Pro Pro Pro Leu His Asn Pro Asn His Ile Phe Ala Arg His Phe
195 200 205
Ser Glu Asn Gly Ala Ala Val Leu Gln Ala Asp Gly Val Leu Val Asn
210 215 220
Thr Phe Glu Ala Leu Glu Gly Asp Ile Leu Ser Ala Leu Asn Ala Gly
225 230 235 240
Thr Val Leu Pro Gly Leu Pro Pro Val Ile Ala Ile Gly Pro Gln Glu
245 250 255
Pro Leu Lys Leu Gly Ala Ser Ala Arg Pro Ala Gly Glu Pro Asp Gly
260 265 270
Trp Ala Thr Trp Leu Asp Ala Gln Pro Pro Lys Ser Val Leu Tyr Val
275 280 285
Ala Phe Gly Asn Arg Thr Ala Leu Ser Thr Val Gln Ile Arg Glu Leu
290 295 300
Gly Lys Gly Leu Glu Lys Ser Gly Cys Arg Phe Leu Trp Val Val Lys
305 310 315 320
Ser Lys Ile Val Asp Lys Asp Glu Ser Val Glu Leu Asp Glu Leu Leu
325 330 335
Gly Glu Gly Tyr Leu Glu Arg Val Gly Asp Arg Gly Met Val Ala Lys
340 345 350
Ser Trp Val Asp Gln Glu Glu Val Leu Ala His Pro Ser Val Gly Gly
355 360 365
Phe Leu Ser His Gly Gly Cys Ser Ser Ser Gln Glu Ala Ile Leu His
370 375 380
Gly Val Arg Ser Leu Asn Trp Pro Gly Met Gly Asp Gln Arg Met Val
385 390 395 400
Ala Ala Gln Ala Glu Lys Arg Gly Ala Gly Val Trp Val Lys Glu Trp
405 410 415
Gly Trp Ser Ser Arg Val His Arg Glu Glu Pro Glu Pro Val Ser Ser
420 425 430
Glu Glu Ile Ala Lys Arg Val Lys Glu Leu Met Glu Asp Asp Ala Val
435 440 445
Gly Glu Ala Met Arg Arg Ile His Glu Glu Ala Val Met Gly Leu Arg
450 455 460
Pro Gly Gly Ser Ser Tyr Glu Gly Leu Arg Lys Phe Val Ala Leu Val
465 470 475 480
Glu Ala Lys Arg Arg Ser Ser Pro Pro Gln Val Ser Glu Thr
485 490
<210> 25
<211> 483
<212> PRT
<213> 大薸 (Pistia stratiotes)
<400> 25
Met Ala Arg Glu Pro Ser Gln Leu Val Arg Asp Gly Asp Leu Ile Pro
1 5 10 15
Thr Pro Thr Pro His Ile Val Ile Leu Pro Ser Ala Gly Met Gly His
20 25 30
Leu Thr Val Ser Thr Arg Leu Ala Ala Ala Leu Val Ser His Gly Cys
35 40 45
Leu Val Thr Leu Ile Thr Pro Leu Pro Met Val Ser Glu Ala Glu Ser
50 55 60
Asn Cys Ile Ser Asn Leu Cys Ser Ser Phe Pro His Ile Arg Leu Leu
65 70 75 80
Glu Leu His Leu Leu Pro Phe Asp Ala Pro Asp Val Asp Pro Phe Leu
85 90 95
Arg Gln Phe Glu Ser Ile Arg Arg Ser Ala Ala Leu Leu Glu Pro Leu
100 105 110
Leu Val Ser Val Thr Pro Pro Val Ser Ala Leu Leu Thr Asp Val Thr
115 120 125
Val Phe Ser Ser Val Val Pro Val Thr Lys Ala Leu Arg Ile Pro Ile
130 135 140
Tyr Met Ile Phe Thr Thr Cys Ala Ser Met Leu Ala Leu Val Ser Thr
145 150 155 160
Phe Pro Gly Ile Arg Lys Glu Ala Gly Asp Gly Pro Ile Glu Ser Ile
165 170 175
Asp Ile Pro Gly Val Gly Thr Ile Pro Gly Ser Arg Ile Pro Arg Pro
180 185 190
Leu His Asp Pro Lys His Leu Phe Thr Ile Gln Phe Val Glu Thr Gly
195 200 205
Glu Ala Met Leu Gln Ala Asp Gly Ile Leu Ser Asn Thr Phe Glu Ala
210 215 220
Leu Asp Gly Asp Ile Leu Ala Ala Leu Asn Ala Gly Ala Val Val Pro
225 230 235 240
Gly Phe Pro Pro Val Ile Ala Val Gly Pro Gln Glu Pro Ile Lys Leu
245 250 255
Arg Ala Ala Ala Arg Pro Ala Val Glu Arg Asp Asp Trp Met Ala Trp
260 265 270
Leu Asp Ala Gln Pro Pro Lys Ser Val Leu Tyr Val Ala Phe Gly Asn
275 280 285
Arg Thr Ala Gln Ser Arg Glu Gln Ile Lys Glu Leu Gly Ile Gly Leu
290 295 300
Glu Lys Ser Gly Cys Arg Phe Leu Trp Val Val Lys Ser Lys Ile Val
305 310 315 320
Asp Lys Asp Asp Asp Asp Gly Val Glu Leu Gly Glu Leu Leu Gly Glu
325 330 335
Gly Tyr Leu Glu Arg Val Gly Glu Arg Gly Val Val Val Lys Ser Trp
340 345 350
Val Ile Gln Glu Glu Val Leu Ala His Pro Ser Val Gly Gly Tyr Leu
355 360 365
Ser His Gly Gly Gly Asn Ser Thr Phe Glu Ala Val Leu Tyr Gly Val
370 375 380
Pro Pro Leu Ile Trp Pro Gly Gly Gly Asp Gln Gln Met Val Ala Ala
385 390 395 400
Gln Val Glu Lys Arg Gly Val Gly Val Trp Ile Lys Glu Trp Gly Leu
405 410 415
Cys Gly Val Asn Gln Val Ser Ser Asp Asp Val Ala Lys Arg Val Arg
420 425 430
Glu Met Met Glu Gly Asp Asn Ala Val Gly Glu Ser Val Arg Arg Met
435 440 445
Gln Glu Glu Ala Val Arg Gly Leu Gly Pro Gly Gly Ser Ser Tyr Glu
450 455 460
Gly Leu Arg Lys Phe Ile Ala Gly Val Val Glu Arg Gly Ser Ser Pro
465 470 475 480
Pro Glu Ile
<210> 26
<211> 483
<212> PRT
<213> 玉米 (Zea mays)
<400> 26
Met Ala Pro Pro Ala Thr Asn Ser Pro Gly Glu Val Asp Ser Gly Gly
1 5 10 15
Arg Pro His Val Met Phe Ile Pro Ser Ala Gly Met Gly His Leu Ile
20 25 30
Pro Phe Phe Arg Phe Ile Ala Ala Leu Ala Gly Gln Ser Val Asp Ile
35 40 45
Ser Val Val Thr Val Leu Pro Thr Val Ser Ala Ala Glu Glu Asp Tyr
50 55 60
Phe Thr Ser Leu Phe Ala Ala Leu Pro Arg Val Arg Arg Val Asp Leu
65 70 75 80
His Leu Leu Pro Phe Asp Ala Ser Lys Leu Pro Ser Asn Asp Arg Asp
85 90 95
Pro Phe Val Leu Arg Trp Glu Ala Leu Arg Arg Ser Ala His Leu Leu
100 105 110
Gly Pro Leu Ile Ala Gly Ala Ala Pro Arg Val Ser Ala Val Ile Thr
115 120 125
Asp Val Thr Leu Thr Ser His Val Ile Pro Ile Ala Lys Glu Leu Gly
130 135 140
Val Gln Cys His Val Leu Phe Pro Ser Ser Ala Thr Met Leu Ser Leu
145 150 155 160
Leu Ala Tyr Thr Pro Val His Ile Asp Lys Lys Ala Glu Gln Gly Ser
165 170 175
Asp Ile Gly Asp Val Asp Ile Pro Gly Val Cys Arg Ile Pro Gln Ser
180 185 190
Cys Leu Pro Gln Val Leu Leu His Leu Asp Lys Leu Phe Thr Lys Gln
195 200 205
Phe Ile Ala Asn Gly Arg Glu Ile Ile Asn Ala Asp Gly Phe Leu Val
210 215 220
Asn Thr Phe Asp Ala Leu Glu Pro Val Ala Leu Ala Ala Leu Arg Asp
225 230 235 240
Gly Lys Val Val Pro Gly Phe Pro Pro Val Tyr Ala Ile Gly Pro Leu
245 250 255
Met Ser Gln Gln Asn Ser Val Asp Glu Gly Gly Lys Glu Glu Gly Ser
260 265 270
Gly Ser Pro Val Ala Trp Leu Asp Glu Gln Pro Ala Arg Ser Val Val
275 280 285
Tyr Val Ala Phe Gly Asn Arg Cys Ala Val Ser His Asp Gln Ile Arg
290 295 300
Glu Ile Ala Ala Gly Leu Glu Ala Ser Asn Cys Arg Phe Leu Trp Val
305 310 315 320
Leu Lys Thr Thr Thr Val Asp Arg Asp Asp Ser Ala Val Leu Thr Glu
325 330 335
Leu Leu Gly Glu Glu Phe Leu Glu Arg Val Gln Gly Arg Gly Leu Val
340 345 350
Thr Lys Ala Trp Val Asp Gln Glu Ala Leu Leu Lys His Pro Ala Met
355 360 365
Gly Leu Phe Leu Ser His Ser Gly Trp Asn Ser Val Thr Glu Ala Ala
370 375 380
Ala Ala Ser Val Pro Leu Leu Ala Trp Pro Arg Gly Gly Asp His Arg
385 390 395 400
Val Asn Ala Met Val Thr Val Ser Gly Gly Val Gly Met Trp Met Glu
405 410 415
His Trp Ser Trp Asp Gly Glu Asp Trp Leu Val Thr Gly Val Glu Ile
420 425 430
Ala Lys Lys Val Lys Glu Val Met Ser Asp Glu Ala Val Arg Ala Arg
435 440 445
Thr Thr Arg Thr Ala Glu Glu Ala Ala Lys Ala Val Ala Glu Gly Gly
450 455 460
Thr Ser Tyr Arg Ser Met Glu Glu Phe Ile Ser Ser Val Lys Ala Thr
465 470 475 480
Val Ala Ser
<210> 27
<211> 47
<212> DNA
<213> 人工序列
<400> 27
gggtcgcgga tccgaattcg agctcatggc aagcaccacc aaatcgg 47
<210> 28
<211> 51
<212> DNA
<213> 人工序列
<400> 28
cgagtgcggc cgcaagcttg tcgactgagt tttgtgtgat attgttgaag g 51
<210> 29
<211> 50
<212> DNA
<213> 人工序列
<400> 29
gggtcgcgga tccgaattcg agctcatgag caaatcggaa aatgcagggc 50
<210> 30
<211> 50
<212> DNA
<213> 人工序列
<400> 30
cgagtgcggc cgcaagcttg tcgaccttga tcttcaacga atgaattagg 50
<210> 31
<211> 49
<212> DNA
<213> 人工序列
<400> 31
gggtcgcgga tccgaattcg agctcatggc accgcccacg gtgctcaac 49
<210> 32
<211> 50
<212> DNA
<213> 人工序列
<400> 32
cgagtgcggc cgcaagcttg tcgacagcag ccttgagctt tgcaacaaac 50
<210> 33
<211> 49
<212> DNA
<213> 人工序列
<400> 33
gggtcgcgga tccgaattcg agctcatggc accaccgcct gcggtgctc 49
<210> 34
<211> 50
<212> DNA
<213> 人工序列
<400> 34
cgagtgcggc cgcaagcttg tcgacagttg cagccttgag cttggcgacg 50
<210> 35
<211> 42
<212> DNA
<213> 人工序列
<400> 35
ggacagcaaa tgggtcgccg gatgggggaa aacaacatgt cc 42
<210> 36
<211> 42
<212> DNA
<213> 人工序列
<400> 36
cgacggagct cgaattccgg aacattcttc ttccaaactt cc 42
<210> 37
<211> 41
<212> DNA
<213> 人工序列
<400> 37
ggacagcaaa tgggtcgccg gatgacttcc tgtactaatt c 41
<210> 38
<211> 41
<212> DNA
<213> 人工序列
<400> 38
cgacggagct cgaattcggg atgccttgaa caatcctctt c 41
<210> 39
<211> 42
<212> DNA
<213> 人工序列
<400> 39
ggtggacagc aaatgggtcg ccggatgtct cgggaacagc ag 42
<210> 40
<211> 37
<212> DNA
<213> 人工序列
<400> 40
cttgtcgacg gagctcgaat tcggtgtggt gtatccc 37
<210> 41
<211> 43
<212> DNA
<213> 人工序列
<400> 41
ggtggacagc aaatgggtcg ccggatgtcg ccgccgacgg ctg 43
<210> 42
<211> 42
<212> DNA
<213> 人工序列
<400> 42
cttgtcgacg gagctcgaat tcggtggccg acgcatacca tc 42
<210> 43
<211> 48
<212> DNA
<213> 人工序列
<400> 43
ggtggacagc aaatgggtcg ccggatgaga ccggggagct cggagagg 48
<210> 44
<211> 48
<212> DNA
<213> 人工序列
<400> 44
gcttgtcgac ggagctcgaa ttcggatctc ccgaacacgt gaaccgcg 48
<210> 45
<211> 46
<212> DNA
<213> 人工序列
<400> 45
ggtggacagc aaatgggtcg ccggatgtct cctcctgcgc ctgcgg 46
<210> 46
<211> 46
<212> DNA
<213> 人工序列
<400> 46
gtcgacggag ctcgaattcg ggacgcggtt gatcctcgcg ataaag 46
<210> 47
<211> 43
<212> DNA
<213> 人工序列
<400> 47
ggtggacagc aaatgggtcg ccggatggtt caagagacat cac 43
<210> 48
<211> 43
<212> DNA
<213> 人工序列
<400> 48
cttgtcgacg gagctcgaat tcggggtctc tgaaacctgc ggt 43
<210> 49
<211> 46
<212> DNA
<213> 人工序列
<400> 49
ggtggacagc aaatgggtcg ccggatggcc cgagagccat cccagc 46
<210> 50
<211> 50
<212> DNA
<213> 人工序列
<400> 50
gcttgtcgac ggagctcgaa ttcgggattt ccggtggtga tgaccccctc 50
<210> 51
<211> 39
<212> DNA
<213> 人工序列
<400> 51
ggacagcaaa tgggtcgccg gatggccccg ccggcaacg 39
<210> 52
<211> 43
<212> DNA
<213> 人工序列
<400> 52
gtcgacggag ctcgaattcg gagacgccac ggttgctttg acg 43
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