CN114207108A - 产生糖基化大麻素的基因修饰的宿主细胞 - Google Patents

产生糖基化大麻素的基因修饰的宿主细胞 Download PDF

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CN114207108A
CN114207108A CN202080054246.0A CN202080054246A CN114207108A CN 114207108 A CN114207108 A CN 114207108A CN 202080054246 A CN202080054246 A CN 202080054246A CN 114207108 A CN114207108 A CN 114207108A
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cannabinoid
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尼古拉斯·斯图尔特·威廉·米尔恩
卡米拉·克努森·巴登
内塔吉·珍妮沙瓦里·加拉格
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Abstract

本发明涉及一种基因修饰以在细胞内产生大麻素糖苷的微生物宿主细胞,所述细胞表达编码糖基转移酶的异源基因,所述糖基转移酶与SEQ ID NO:157或207中包括的糖基转移酶具有至少70%同一性,能够使大麻素受体与糖基供体在细胞内糖基化,从而产生所述大麻素糖苷。

Description

产生糖基化大麻素的基因修饰的宿主细胞
技术领域
本发明涉及在细胞内产生大麻素糖苷的基因修饰的宿主细胞;用于这样的宿主细胞的重组多核苷酸构建体和载体,用于这样的宿主细胞的细胞培养物;用于产生大麻素糖苷的方法,由这样的方法产生的发酵液;包括这样的发酵液的组合物和制剂;以及这样的组合物和制剂的用途。
背景技术
数千年来,源自植物诸如大麻(Cannabis Sativa)的大麻素因其药用特性而被消耗。已从植物中分离出超过100种大麻素分子,其中许多与多种人疾病病症相关。最近,大麻素,并且特别是大麻二酚(CBD)和Δ-9-四氢大麻酚(THC)已被批准用作多种病症的治疗药物。CBD和THC是研究最充分的大麻素,可能是因为它们是植物中发现的最丰富的大麻素的事实。
虽然大麻素被认为有希望用于治疗性治疗,但存在若干种特性使大多数大麻素作为治疗分子的用处不大。大麻素是高亲脂性的,具有低生物利用度,并且可迅速从体内清除。此外,一些大麻素,特别是THC,是具精神活性的,这意指它们可能必须以次优剂量给药,以避免引发严重副作用。此外,大麻素在化学上也不稳定,并且即使在环境条件下也迅速降解。因此,这种非期望的特性正在限制大麻素的治疗潜力并阻碍有效治疗的发展。因此,需要改进大麻素的药代动力学和/或治疗特性。WO2017053574提出通过在存在糖基转移酶的情况下将大麻素苷元与糖供体一起孵育来制备大麻素糖苷前药。WO2019014395建议在酵母细胞培养悬浮液中表达糖基转移酶,以及然后将大麻素引入悬浮液中以产生水溶性大麻素。
大麻素的植物原位(in planta)产生需要植物细胞协同进行大量不同的酶介导的化学反应(途径),并且虽然原则上理解植物酶多肽和编码它们的多核苷酸有助于植物中合成大麻素,但大麻素途径的许多方面还有待探索,不仅哪些多肽与自然界中产生特定大麻素有关,而且哪些多肽/酶可以在植物外(例如在异源宿主细胞中)产生大麻素,并且特别是当通过植物外生物合成制备方法产生时,哪些多肽/酶能够产生更高产量的期望的大麻素。
因此,仍需要具有改进的药代动力学和/或治疗特性的大麻素以及有效产生这种改进的大麻素的方法。
发明内容
本发明的发明人已发现糖基转移酶,其不仅出人意料地整合并起作用以在基因修饰的宿主细胞中的细胞内产生大麻素糖苷,而且在产生大麻素糖苷方面比迄今为止已知的方法表现出显著改进。因此,在第一方面,本发明提供了一种经基因修饰以在细胞内产生大麻素糖苷的微生物宿主细胞,所述细胞表达编码至少一种糖基转移酶的异源基因,所述糖基转移酶能够使大麻素受体与糖基或糖供体发生细胞内糖基化,从而产生大麻素糖苷。
在另外的方面,本发明提供了一种包括编码本发明的糖基转移酶的多核苷酸序列的多核苷酸构建体,其可操作地连接到与糖基编码多核苷酸异源的一个或多个控制序列。
在另外的方面,本发明提供了一种包括本发明的多核苷酸构建体的表达载体。
在另外的方面,本发明提供了一种包括本发明的多核苷酸构建体或载体的基因修饰的宿主细胞。
在另外的方面,本发明提供了一种包括本发明的基因修饰的宿主细胞和生长培养基的细胞培养物。
在另外的方面,本发明提供了一种用于产生大麻素糖苷的方法,包括:
a)在允许基因修饰的宿主细胞产生所述大麻素糖苷的条件下培养本发明的细胞培养物;以及
b)任选地回收和/或分离所述大麻素糖苷。
在另外的方面,本发明提供了一种包括本发明的细胞培养物中包括的所述大麻素糖苷的发酵液。
在另外的方面,本发明提供了一种组合物,所述组合物包括本发明的发酵液或大麻素糖苷以及一种或多种剂、添加剂和/或赋形剂。
在另外的方面,本发明提供了一种大麻素糖苷,包括与选自以下的糖共价连接的大麻素苷元或大麻素糖苷:木糖;鼠李糖;半乳糖;N-乙酰葡糖胺;N-乙酰半乳糖胺;和阿拉伯糖,或者包括通过1,4-或1,6-糖苷键共价键连接到糖苷部分的大麻素苷元或大麻素糖苷。
在另外的方面,本发明提供了一种用于制备药物制剂的方法,所述方法包括将本发明的组合物与一种或多种药物级赋形剂、添加剂和/或佐剂混合。
在另外的方面,本发明提供了一种可获自用于制备药物制剂的本发明的方法的药物制剂。
在另外的方面,本发明提供了一种可获自用于制备药物制剂的本发明的方法的药物制剂,用作药物用途。
在另外的方面,本发明提供了一种用于治疗哺乳动物中疾病的方法,包括向哺乳动物给药治疗有效量的本发明的药物制剂。
附图说明
图1示出了从葡萄糖产生大麻素的微生物途径。
图2示出了展示酿酒酵母中多个整合片段的体内同源重组的示意图。
图3示出了由在酿酒酵母中引入实施例17中描述的质粒引起的用于产生大麻素和大麻素糖苷的生物合成途径。
图4示出了通过LC-MS-QTOF验证的大麻素糖苷的结构。
图5示出了来自通过Cs73Y将CBG体外转化为CBG糖苷的 LC-MS-QTOF色谱图的实例。
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具体实施方式
定义
如本文所用,术语“ACT”是指能够将两分子乙酰-CoA转化为乙酰乙酰 -CoA的乙酰乙酰-CoA硫解酶(EC 2.3.1.9)。ACT也称为ERG10。
如本文所用,术语“HCS”是指能够将乙酰乙酰-CoA和乙酰-CoA转化为 HMG-CoA的羟甲基戊二酰基-CoA(HMG-CoA)合酶酶(EC 4.1.3.5)。HCS也称为ERG13。
如本文所用,术语“HCR”是指能够将HMG-CoA转化为甲羟戊酸的 HMG-CoA还原酶(EC1.1.1.34)。
如本文所用,术语“MVK”是指能够将甲羟戊酸转化为甲羟戊酸-5-磷酸的甲羟戊酸激酶(EC2.7.1.36)。MVK也称为ERG12。
如本文所用,术语“PMK”是指能够将甲羟戊酸-5-磷酸转化为甲羟戊酸二磷酸的磷酸甲羟戊酸激酶(EC2.7.4.2)。PMK也称为ERG8。
如本文所用,术语“MPC”是指能够将甲羟戊酸二磷酸转化为异戊烯基二磷酸(IPP)的甲羟戊酸焦磷酸脱羧酶(EC4.1.1.33)。MPC也称为MVD1。
如本文所用,术语“IPI”是指能够将IPP转化为二甲基烯丙基二磷酸 (DMAPP)的异戊烯基二磷酸异构酶(EC5.3.3.2)。IPI也称为IDI1。
如本文所用,术语“GPPS”是指能够将DMAPP和IPP转化为香叶基二磷酸(GPP)的香叶基二磷酸合酶(EC2.5.1.1)。
如本文所用,术语“AAE”是指能够将乙酰-CoA和己酸或乙酰-CoA和丁酸分别转化为己酰基-CoA或丁酰基-CoA的酰基活化酶(EC6.2.1.2)。
如本文所用,术语“TKS”是指能够将己酰基-CoA和丙二酰基-CoA或丁酰基-CoA和丙二酰基-CoA分别转化为3,5,7-三氧亚基十二烷酰基-CoA或 3,5,7-三氧亚基十一烷酰基-CoA的3,5,7-三氧亚基十二烷酰基-CoA合酶(EC2.3.1.206)。TKS也称为橄榄醇合酶。
如本文所用,术语“OAC”是指能够分别将3,5,7-三氧亚基十二烷酰基 -CoA转化为橄榄酸或将3,5,7-三氧亚基十一烷酰基-CoA转化为divarinolic acid的3,5,7-三氧亚基十二烷酰基-CoA环化酶或3,5,7-三氧亚基十一烷酰基 -CoA环化酶(EC4.4.1.26)。OAC也称为橄榄酸环化酶。
如本文所用,术语“CBGAS”是指能够将GPP和橄榄酸(OA)或GPP和 divarinolicacid(DVA)分别转化为大麻萜酚酸(CBGA)或次大麻萜酚酸 (CBGVA)的大麻萜酚酸合酶(2.5.1.102)。
如本文所用,术语“CBDAS”是指能够将CBGA或CBGVA分别转化为大麻二酚酸(CBDA)或次大麻二酚酸(CBDVA)的大麻二酚酸合酶 (EC1.21.3.8)。
如本文所用,术语“THCAS”是指能够将CBGA或CBGVA分别转化为四氢大麻酚酸(tetrahydrocannabinolic acid)(THCA)或四氢次大麻酚酸 (tetrahydrocannabivarinicacid)(THCVA)的四氢大麻酚酸合酶(EC1.21.3.7)。
如本文所用,术语“CBCAS”是指能够将CBGA或CBGVA分别转化为大麻色烯酸(CBCA)或annabichromevarinic acid的大麻色烯酸合酶 (EC1.21.99.-或EC1.3.3.-)。
如本文所用,术语“糖基转移酶”或“GT”是指通过将糖基基团(糖)从活化的糖基供体转移到亲核糖基受体分子(其亲核体可以是且尤其是基于氧、碳、氮或硫的)来催化糖苷形成的酶(EC2.4)。糖基转移的产物可以是O-、N-、 S-或C-糖苷。在本发明的背景下,亲核糖基受体是大麻素或大麻素糖苷,并且糖基转移的产物是O-或C-糖苷。
如本文所用,关于糖基供体的术语“核苷酸糖苷”是指包括与糖基基团共价连接的核苷酸部分的化合物,其中核苷酸包括与一个或多个磷酸基团共价连接的核苷。这样的化合物也称为“活化糖苷”,并且其中糖基基团是糖,称为“核苷酸糖”或“活化糖”。
如本文所用,术语“异源的”或“重组的”及其语法等同物是指“源自不同物种或细胞”的实体。例如,异源或重组多核苷酸基因是宿主细胞中不是天然含有该基因的基因,即该基因来自与宿主细胞不同的物种或细胞类型。
如本文所用,术语“基因修饰的宿主细胞”是指包括并表达异源或重组多核苷酸基因的宿主细胞。
如本文所用,术语“底物”或“前体”是指可以转化为不同化合物的任何化合物。例如,IPP可以是将IPP转化为DMAPP的IPI的底物。为了简明,底物和/或前体包括通过细胞中的酶促反应原位产生的化合物或外源提供的化合物(例如宿主细胞可代谢成期望化合物的外源提供的有机碳分子)两者。
如本文所用,术语“代谢途径”旨在一指两种或更多种酶在活细胞中的反应链中起作用(依次或被中间步骤中断)以将一种或多种化学底物转化为一种或多种化学产物。酶的特点是具有催化活性,其可以改变一种或多种底物的化学结构。酶可以具有超过一种底物并产生超过一种产物。酶还可以依赖于辅助因子,该辅助因子可以是无机化合物或有机化合物,诸如蛋白质,例如酶(辅酶)。NADPH和NAD+是辅因子的实例。
术语“起作用的生物合成代谢途径”是指在活的重组宿主中发生的代谢途径,如本文所述。
如本文所用,术语“体内”是指在活细胞(包括,例如,微生物或植物细胞(植物原位))内。
如本文所用,术语“体外”是指在活细胞外,包括但不限于,例如,在微孔板、管、烧瓶、烧杯、罐、反应器等中。
如本文所用,术语“基本上”或“大约”或“约”是指围绕值或参数的合理偏差,使得该值或参数没有显著改变。值的偏差的这些术语应被解释为包括该值的偏差,其中该偏差将不否定偏离该值的含义。例如,关于参考数值,程度的术语可以包括该值加或减10%的值范围。例如,使用这些偏差术语还可以包括指定值加或减,诸如加或减9%、8%、7%、6%、5%、4%、3%、 2%或1%的偏差范围。
如本文所用,术语“和/或”旨在意指表示包括性的“或”。措辞X和/或Y 旨在意指X或Y以及X和Y。此外,措辞X、Y和/或Z旨在表示单独的X、 Y和Z,或X、Y和Z的任何组合。
如本文所用,关于化合物的术语“分离的”或“纯化的”或“提取的”或“回收的”可互换使用,是指通过人为干预,已被置于与其在自然界中被发现时所处的形式或环境不同的形式或环境中的任何化合物。分离的化合物包括,但不限于本发明的化合物,其中该化合物相对于它们在自然界中与之相关的其他成分的比例增加或减少。在重要的实施方式中,化合物的量相对于在自然界中与该化合物相关的其他成分增加。在实施方式中,本发明的化合物可以被分离成纯的或基本上纯的形式。在该背景下,基本上纯的化合物意指该化合物与从产生该化合物开始时就存在的或在制备工艺中生成的其他外源性或非期望的物质分离。这样的基本上纯的化合物制剂包含按重量计小于10%,诸如小于8%、诸如小于6%、诸如小于5%、诸如小于4%、诸如小于3%、诸如小于2%、诸如小于1%、诸如小于0.5%的其他外源性或非期望的物质,当天然地或重组表达化合物时,该其他外源性或非期望的物质通常与该化合物相关。在实施方式中,分离的化合物按重量计为至少90%纯的,诸如至少91%纯的、诸如至少92%纯的、诸如至少93%纯的、诸如至少94%纯的、诸如至少95%纯的、诸如至少96%纯的、诸如至少97%纯的、诸如至少98%纯的、诸如至少99%纯的、诸如至少99.5%纯的、诸如100%纯的。
如本文所用,关于物质的术语“非天然存在的”是指通常未在自然界或天然生物系统中发现的任何物质。在该背景下,“在自然界或天然生物系统中发现”这一术语不包括通过有意或意外的人为干预将物质释放至自然界而在自然界中发现物质。非天然存在的物质可以包括完全或部分通过人为干预合成的物质和/或通过人为修饰天然物质制备的物质。
术语“同一性%”在本文中用于关于两个氨基酸序列之间或两个核苷酸序列之间的相关性。如本文所用,关于氨基酸序列的“同一性%”是指当使用在EMBOSS包(EMBOSS:欧洲分子生物学开放软件套件,Rice等人,2000,遗传学趋势(Trends Genet.)16:276-277(优选为5.0.0版本或更高版本)的 Needle程序中执行的Needleman-Wunsch算法(Needleman和Wunsch,1970,分子生物学杂志(Mol.Biol.)48:443-453时获得的两个氨基酸序列之间以百分比表示的同一性程度。使用的参数是空位开放罚分为10,空位延伸罚分为0.5,以及EBLOSUM62(BLOSUM62的EMBOSS版本)替换矩阵。标记为“最长同一性”的Needle输出(使用-nobrief选项获得)用作同一性百分比并计算如下:
Figure BDA0003490979370000081
如本文所用,关于核苷酸序列的“同一性%”是指当使用在EMBOSS包 (EMBOSS:欧洲分子生物学开放软件套件,Rice等人,2000,同上)(优选为5.0.0版本或更高版本)的Needle程序中执行的Needleman-Wunsch算法 (Needleman和Wunsch,1970,同上)时获得的两个核苷酸序列之间以百分比表示的同一性程度。使用的参数是空位开放罚分为10,空位延伸罚分为 0.5,以及EDNAFULL(NCBI NUC4.4的EMBOSS版本)替换矩阵。标记为“最长同一性”的Needle输出(使用-nobrief选项获得)用作同一性百分比并计算如下:
Figure BDA0003490979370000082
本发明的蛋白质序列可以进一步用作“查询序列”以对序列数据库进行搜索,例如以识别其他家族成员或相关序列。可以使用BLAST程序进行这样的搜索。进行BLAST分析的软件可以通过国家生物技术信息中心公开获得(http://www.ncbi.nlm.nih.gov)。BLASTP用于氨基酸序列,以及BLASTN 用于核苷酸序列。BLAST程序使用默认:
-空位开放罚分:默认=5(用于核苷酸)/11(用于蛋白质)
-空位延伸罚分:默认=2(用于核苷酸)/1(用于蛋白质)
-核苷酸错配罚分:默认=-3
-核苷酸匹配加分:默认=1
-期望值:默认=10
-字长:默认=11(用于核苷酸)/28(用于megablast)/3(用于蛋白质)。
此外,氨基酸序列查询或核酸序列查询与检索到的同源序列之间的局部同一性程度通过BLAST程序确定。然而,仅那些给出高于某些阈值的匹配的序列区段被比较。因此,程序仅计算这些匹配区段的同一性。因此,以这种方式计算的同一性称为局部同一性。
术语“cDNA”是指可以通过从真核或原核细胞获得的成熟、剪接的 mRNA分子逆转录制备的DNA分子。cDNA缺少可能存在于相应基因组 DNA中的内含子序列。最初的初级RNA转录物是mRNA的前体,其通过一系列步骤进行加工,包括剪接,然后作为成熟的剪接mRNA出现。
术语“编码序列”是指直接指定多肽的氨基酸序列的核苷酸序列。编码序列的边界通常由开放阅读框确定,其以起始密码子(诸如ATG、GTG或TTG) 开始并以终止密码子(诸如TAA、TAG或TGA)结束。编码序列可以是基因组DNA、cDNA、合成DNA或其组合。
如本文所用,术语“控制序列”是指表达编码多肽的多核苷酸所必需的核苷酸序列。控制序列对于编码多肽的多核苷酸可以是天然的(即,来自相同基因)或者是异源的或外源的(即,来自不同的基因)。控制序列包括,但不限于前导序列、聚腺苷酸化序列、前肽编码序列、启动子序列、信号肽编码序列、翻译终止子(终止)序列和转录终止子(终止)序列。要成为可操作的控制序列,通常必须包括启动子序列、转录和翻译终止信号。为了引入促进控制序列与编码多肽的多核苷酸的编码区连接的特定限制性位点的目的,控制序列可以与接头一起提供。
术语“表达”包括涉及产生多肽的任何步骤,包括但不限于转录、转录后修饰、翻译、翻译后修饰和分泌。
术语“表达载体”是指包括编码多肽的多核苷酸并与提供其表达的控制序列可操作地连接的线性或环状DNA分子。
术语“宿主细胞”是指对用包括本发明多核苷酸的多核苷酸构建体或表达载体进行转化、转染、转导等敏感的任何细胞类型。术语“宿主细胞”涵盖由于复制期间发生的突变而与亲本细胞不相同的亲本细胞的任何后代。
术语“多核苷酸构建体”是指单链或双链多核苷酸,其从天然存在的基因中分离或被修饰以以自然界中不存在的其他方式包含核酸区段,或者其是被合成的,并且其包括一个或多个控制序列。
术语“可操作地连接”是指其中控制序列位于相对于编码多核苷酸的适当位置,使得控制序列指导编码多核苷酸的表达的构型。
术语“核苷酸序列”和“多核苷酸”在本文中可互换使用。
在整个说明书和所附权利要求中使用的术语“包括(comprise)”和“包括(include)”以及变体诸如“包括(comprises、comprising)”和“包括(includes、including)”应被包括性地解释。在背景允许的情况下,这些词旨在传达可能包括未具体列举的其他要素或整数。
冠词“一个(种)(a和an)”在本文中用于指代冠词的一个(种)或多于一个 (种)(即一个(种)或至少一个(种))语法对象。例如,“一个要素”可以指一个要素或多于一个要素。
术语如“优选地”、“普遍地”、“特别地”和“典型地”在本文中不用于限制要求保护的发明的范围或暗示某些特征对于要求保护的发明的结构或功能是关键的、必不可少的,或甚至是重要的。相反,这些术语仅旨在强调可以或不可以在本发明的特定实施方式中使用的替代的或另外的特征。
如本文所用,术语“细胞培养物”是指包括多个本发明的基因修饰的宿主细胞的培养基。细胞培养物可以包括基因修饰的宿主细胞的单一菌株或可以包括基因修饰的宿主细胞的两种或更多种不同的菌株。培养基可以是适用于基因修饰的宿主细胞的任何培养基,例如,液体培养基(即,培养液) 或半固体培养基,并且可以包括另外的组分,例如,碳源,诸如右旋糖、蔗糖、甘油或乙酸盐;氮源,诸如硫酸铵、脲或氨基酸;磷酸盐来源;维生素;微量元素;盐类;氨基酸;核碱基;酵母提取物;氨基糖苷类抗生素,诸如G418和潮霉素B。
术语“1'-O”和“3'-O”是指大麻素上1'和3'位的OH基团。由于包含两个 OH基团(例如CBD、CBDV、CBG)的大麻素的对称性质以及在这些分子中发生的自由旋转,术语“1'-O”和“3'-O”可以互换使用。例如,应理解CBD-1'-O-β-D-木糖苷和CBD-3'-O-β-D-木糖苷可以互换使用以描述相同的分子。
术语“二糖苷”、“三糖苷”和“四糖苷”是指具有以任何O-连接附连在一起的2、3和4个糖苷部分的分子。例如CBD-1'-O-β-D-二-木糖苷是指在CBD 的1'位置附连1个木糖并且在第一木糖的任意位置附连第二木糖的CBD分子。
术语“龙胆二糖苷”、“纤维二糖苷”和“昆布二糖苷”是指其中两个葡萄糖部分分别通过1,6-位置、1,4-位置或1,3-位置处的O-β-糖苷键连接的二糖苷分子。
根据3D结构和反应机制,糖基转移酶可以进一步分为不同的GT家族。更特别地,GT1超家族是指含有结合UDP-糖的PSPG盒的UDP糖基转移酶(UGT)。根据氨基酸同一性,UGT超家族成员可以进一步分为由UGT命名委员会(Mackenzie等人,1997)定义的家族和亚家族。>40%的同一性属于相同的UGT家族,例如UGT73,,并且>60%的氨基酸同一性定义了亚家族,例如UGT73Y
基因修饰的宿主细胞
一方面,本发明提供了一种基因修饰以在细胞内产生大麻素糖苷的微生物宿主细胞,所述细胞表达编码至少一种糖基转移酶的异源基因,该糖基转移酶能够使大麻素受体与糖基供体发生细胞内糖基化,从而产生大麻素糖苷。
大麻素受体
大麻素受体可以是异戊二烯基供体和异戊二烯基受体的缩合产物或其衍生物。大麻素受体可以是大麻素苷元或大麻素糖苷。
异戊二烯基供体可以选自香叶基二磷酸、橙花基二磷酸、法呢基二磷酸、二甲基烯丙基二磷酸和香叶基香叶基焦磷酸的组。特别地,异戊二烯基供体是香叶基二磷酸(GPP)。异戊二烯基受体可以是选自己酸、丁酸、戊酸、庚酸、辛酸、壬酸、癸酸、4-甲基己酸、5-己酸和6-庚酸的组的脂肪酸的衍生物。特别地,异戊二烯基受体选自橄榄酸、divarinolicacid、橄榄醇、呋罗苯异戊酮(phlorisovalerophenone)、白藜芦醇、柚皮素、间苯三酚和尿黑酸的组,并且在一种实施方式中异戊二烯基受体是橄榄酸和/或divarinolic acid。
合适的大麻素受体是其中大麻素受体和/或大麻素糖苷具有充当人或动物大麻素受体的激动剂或拮抗剂的亲和力的那些。人的不同大麻素受体是已知的,包括但不限于CB1、CB2、GPR55、5-HT1A、TRPV1和TRPA1。已知一些大麻素受体具有精神活性,诸如THC,其被认为与大脑中的CB1 受体结合,并通过细胞内活化,诱导机体和大脑中自然产生的阿那达胺 (anandamide)和2-花生四烯酸甘油合成。在一种实施方式中,当例如通过使用可从Eurofins (https://www.eurofinsdiscovery.com/HTS019RTA-Ready-to-Assay-CB1- Cannabinoid-Receptor-Frozen-Cells/)获得的HTS019RTA-READY-TO-ASSAYTM CB1CANNABINOID RECEPTOR FROZEN CELLS进行测定时,大麻素受体是非精神作用的或是比THC低至少25%的精神作用的。优选地,大麻素受体和/或大麻素糖苷比THC低至少50%的非精神作用,诸如比THC低至少75%的精神作用,或比THC低至少80%,或至少90%,或至少95%的精神作用。
大麻素受体通常是中性或酸性的,并且在实施方式中可以选自以下的组:大麻色烯型(CBC)、大麻萜酚型(CBG)、大麻二酚型(CBD)、四氢大麻酚型(THC)、大麻环酚型(CBL)、大麻艾尔松型(CBE)、大麻酚型(CBN)、脱氢大麻二酚型(CBND)和二羟基大麻酚型(CBT)。更特定地,大麻素受体选自以下的组:大麻萜酚酸(CBGA)、大麻萜酚酸单甲基醚(CBGAM)、大麻萜酚单甲基醚(CBGM)、次大麻萜酚酸(CBGVA)、次大麻萜酚(CBGV)、大麻色烯酸(CBCA)、次大麻色烯酸(CBCVA)、次大麻色烯(CBCV)、大麻二酚酸 (CBDA)、大麻二酚单甲基醚(CBDM)、大麻二酚-C4(CBD-C4)、次大麻二酚酸(CBDVA)、次大麻二酚(CBDV)、大麻二酚可尔(CBD-C1)、Δ9-反式四氢大麻酚(Δ9-THC)、Δ9-四氢大麻酚(Δ9-THC)、Δ9-顺式四氢大麻酚(Δ9-THC)、四氢大麻酚酸(THCA)、Δ9-四氢大麻酚酸A(THCA-A)、Δ9-四氢大麻酚酸B(THCA-B)、Δ9-四氢大麻酚酸-C4(THCA-C4)、Δ9-四氢大麻酚-C4(THC-C4)、Δ9-四氢次大麻酚酸(THCVA)、Δ9-四氢次大麻酚(THCV)、Δ9-四氢大麻酚可尔酸(THCA-C1)、Δ9-四氢大麻酚可尔(THC-C1)、Δ7-顺式-异-四氢次大麻酚、Δ8-四氢大麻酚酸(Δ8-THCA)、Δ8-反式-四氢大麻酚(Δ8-THC)、Δ8-四氢大麻酚 (Δ8-THC)、Δ8-顺式-四氢大麻酚(Δ8-THC)、大麻环酚酸(CBLA)、大麻环酚 (CBL)、次大麻环酚(CBLV)、大麻艾尔松酸A(CBEA-A)、大麻艾尔松酸 B(CBEA-B)、大麻艾尔松(CBE)、cannabielsoinic acid、大麻二吡喃环烷、大麻二吡喃环烷酸、大麻酚酸(CBNA)、大麻酚甲基醚(CBNM)、大麻酚 -C4(CBN-C4)、次大麻酚(CBV)、大麻酚-C2(CNB-C2)、大麻酚可尔(CBN-C1)、脱氢大麻二酚(CBND)、脱氢次大麻二酚(CBVD)、二羟基大麻酚(CBT)、10- 乙氧基-9-羟基-δ-6a-四氢大麻酚、8,9-二羟基-δ-6a-四氢大麻酚、二羟基次大麻酚(CBTVE)、脱氢大麻呋喃(DCBF)、大麻呋喃(CBF)、大麻色酮(CBCN)、cannabiciuan(CBT)、10-氧亚基-δ-6a-四氢大麻酚(OTHC)、δ-9-顺式-四氢大麻酚(顺式-THC)、3,4,5,6-四氢-7-羟基-α-α-2-三甲基-9-正丙基-2,6-桥亚甲基 -2H-l-苯并氧杂环辛三烯-5-甲醇(OH-异-HHCV)、大麻利比索(CBR)、三羟基-δ-9-四氢大麻酚(triOH-THC)、perrottetinene、perrottetinenic acid、11-Nor-9- 羧基-THC、11-羟基-Δ9-THC、Nor-9-羧基-Δ9-四氢大麻酚、 tetrahydrocannabiphorol(THCP)、cannabidiphorol(CBDP)、Cannabimovone(CBM)及其衍生物,或所述大麻素受体是选自以下组的内源性大麻素:花生四烯酰乙醇酰胺(花生四烯酰乙醇胺,AEA)、2-花生四烯酰乙醇酰胺(2-AG)、1-花生四烯酰乙醇酰胺(1-AG)和二十二碳六烯酰乙醇酰胺 (DHEA,synaptamide)、油酰乙醇酰胺(OEA)、二十碳五烯酰乙醇酰胺、前列腺素乙醇酰胺、二十二碳六烯酰乙醇酰胺、亚麻酰乙醇酰胺、5(Z),8(Z),11(Z)-二十碳三烯酸乙醇酰胺(米德酸乙醇酰胺)、十七烷醇乙醇酰胺、硬脂酰乙醇酰胺、二十二碳烯酰乙醇酰胺、神经酰基乙醇酰胺、二十三酰乙醇酰胺、木蜡酰乙醇酰胺、肉豆蔻酰乙醇酰胺、十五烷酰乙醇酰胺、棕榈油酰乙醇酰胺、二十二碳六烯酸(DHA)。在另一种实施方式中,大麻素受体是选自以下的组的内源性大麻素:花生四烯酰乙醇酰胺(anandamide, AEA)、2-花生四烯酰乙醇酰胺(2-AG)、1-花生四烯酰乙醇酰胺(1-AG)和二十二碳六烯酰乙醇酰胺(DHEA,synaptamide)、油酰乙醇酰胺(OEA)、二十碳五烯酰乙醇酰胺、前列腺素乙醇酰胺、二十二碳六烯酰乙醇酰胺、亚麻酰乙醇酰胺、5(Z),8(Z),11(Z)-二十碳三烯酸乙醇酰胺(米德酸乙醇酰胺)、十七烷醇乙醇酰胺、硬脂酰乙醇酰胺、二十二碳烯酰乙醇酰胺、神经酰基乙醇酰胺、二十三酰乙醇酰胺、木蜡酰乙醇酰胺、肉豆蔻酰乙醇酰胺、十五烷酰乙醇酰胺、棕榈油酰乙醇酰胺和二十二碳六烯酸(DHA)。其他列于 Elsohly M.A.和Slade D.;生命科学2005;78;第539-548页(Elsohly M.A.and Slade D.;Life Sci.2005;78;pp539 548.)。
酸性大麻素受体可以通过热、光或碱性条件脱羧成它们的中性对应物。
糖基供体
合适的糖基供体是核苷酸糖苷。可用于本发明的核苷酸糖苷包括核苷三磷酸糖苷(NTP-糖苷)、核苷二磷酸糖苷(NDP-糖苷)和核苷单磷酸糖苷 (NMP-糖苷)。糖单或二磷核苷酸(有时称为Leloir供体);以及相应的GT被称为Leloir糖基转移酶。特别优选的核苷是尿苷、腺苷、鸟苷、胞苷和/或脱氧胸苷。有用的核苷酸糖苷包括尿苷二磷酸糖苷(UDP-糖苷)、腺苷二磷酸糖苷(ADP-糖苷)、胞苷二磷酸糖苷(CDP-糖苷)、胞苷单磷酸糖苷(CMP- 糖苷)、脱氧胸苷二磷酸糖苷(dTDP-糖苷)和鸟苷二磷酸糖苷(GDP-糖苷)。
特别有用的UDP-糖基供体是UDP-D-葡萄糖(UDP-Glc);UDP-半乳糖 (UDP-Gal);UDP-D-木糖(UDP-Xyl);UDP-N-乙酰-D-葡糖胺(UDP-GlcNAc); UDP-N-乙酰-D-半乳糖胺(UDP-GalNAc);UDP-D-葡糖醛酸(UDP-GlcA); UDP-L-鼠李糖(UDP-Rham);UDP-D-呋喃半乳糖(UDP-Galf);UDP-阿拉伯糖;UDP-芹菜糖;UDP-2-乙酰胺基-2-脱氧-α-D-甘露糖醛酸酯(mannuronate); UDP-N-乙酰-D-半乳糖胺4-硫酸酯;UDP-N-乙酰-D-甘露糖胺;UDP-2,3-双(3-羟基十四烷酰基)-葡糖胺;UDP-4-脱氧-4-甲酰胺基-β-L-阿拉伯吡喃糖; UDP-2,4-双(乙酰胺基)-2,4,6-三脱氧-α-D-吡喃葡萄糖;UDP-半乳糖醛酸酯和 /或UDP-3-氨基-3-脱氧-α-D-葡萄糖。其他有用的核苷酸糖苷糖基供体是鸟苷二磷酸-D-甘露糖(GDP-Man);鸟苷二磷酸-L-岩藻糖(GDP-Fuc);鸟苷二磷酸-L-鼠李糖(GDP-Rha);胞苷单磷酸-N-乙酰神经氨酸(CMP-Neu5Ac);胞苷单磷酸-2-酮-3-脱氧-D-甘露辛酸(CMP-Kdo)。腺苷二磷酸糖(ADP-糖),诸如ADP-Glc,也可用作糖基供体。特别地,供体是UDP并且GT是UDP 依赖性糖基转移酶(UGT)。
糖基转移酶
本发明的糖基转移酶可以源自真核、原核或古生物来源。在一个实施方式中,来源是真核生物,诸如哺乳动物(例如人)、植物或真菌。有用的植物包括但不限于水稻(Oryzasativa)、番红花(Crocus sativus)、烟草(Nicotiana tabacum)、甜叶菊(Steviarebaudiana)、本氏烟草(Nicotiana benthamiana)和拟南芥(Arabidopsis thaliana)。此外,糖基转移酶可以能够使用核苷酸糖苷(诸如NTP-糖苷、NDP-糖苷和/或NMP-糖苷)作为糖基供体使大麻素糖基化。特别地,能够使用核苷酸糖苷的糖基转移酶是有用的,其中核苷选自尿苷、腺苷、鸟苷、胞苷和脱氧胸苷作为糖基供体。在另外的实施方式中,糖基转移酶可以使用糖基供体使大麻素糖基化,该糖基供体选自UDP-糖苷、 ADP-糖苷、CDP-糖苷、CMP-糖苷、dTDP-糖苷和GDP-糖苷。特别地,UDP- 和/或ADP-糖基转移酶是有用的。
进一步有用的糖基转移酶是可以使用选自以下的一种或多种糖基供体糖基化大麻素生物的那些糖基转移酶:UDP-D-葡萄糖(UDP-Glc);UDP-D- 半乳糖(UDP-Gal);UDP-D-木糖(UDP-Xyl);UDP-L-鼠李糖(UDP-Rham); UDP-N-乙酰-D-葡糖胺(UDP-GlcNAc);UDP-N-乙酰-D-半乳糖胺 (UDP-GalNAc);UDP-D-葡糖醛酸(UDP-GlcA);UDP-D-呋喃半乳糖 (UDP-Galf);UDP-L-阿拉伯糖;UDP-D-芹菜糖;UDP-2-乙酰胺基-2-脱氧 -α-D-甘露糖醛酸酯(mannuronate);UDP-N-乙酰-D-半乳糖胺4-硫酸酯; UDP-N-乙酰-D-甘露糖胺;UDP-2,3-双(3-羟基十四烷酰基)-葡糖胺;UDP-4- 脱氧-4-甲酰胺基-β-L-阿拉伯吡喃糖;UDP-2,4-双(乙酰胺基)-2,4,6-三脱氧 -α-D-吡喃葡萄糖;UDP-半乳糖醛酸酯和UDP-3-氨基-3-脱氧-α-D-葡萄糖。其他有用的糖基供体是鸟苷二磷酸-D-甘露糖(GDP-Man);鸟苷二磷酸-L-岩藻糖(GDP-Fuc);鸟苷二磷酸-L-鼠李糖(GDP-Rha);胞苷单磷酸-N-乙酰神经氨酸(CMP-Neu5Ac);胞苷单磷酸-2-酮-3-脱氧-D-甘露辛酸(CMP-Kdo)。
其他有用的糖基转移酶是大麻素苷元O-糖基转移酶;大麻素糖苷O- 糖基转移酶;大麻素苷元O-葡糖基转移酶;大麻素苷元O-鼠李糖基转移酶;大麻素苷元O-木糖基转移酶;大麻素苷元O-阿拉伯糖基转移酶;大麻素苷元O-N-乙酰氨基半乳糖基转移酶;大麻素苷元O-N-乙酰氨基葡糖基转移酶;大麻素苷元/糖苷单-O-糖基转移酶;大麻素苷元/糖苷二-O-糖基转移酶;大麻素苷元/糖苷三-O-糖基转移酶;大麻素苷元/糖苷四-O-糖基转移酶;大麻素O-半乳糖基转移酶和/或大麻素O-葡糖醛酸基转移酶。
又另外使用的糖基转移酶是O-糖苷转移酶和/或C-糖苷转移酶。有用的糖基转移酶可以属于酶分类EC2.4.1.-或EC2.4.2.-。来自EC2.4.1.-的糖基转移酶,诸如来自EC2.4.1.17(使用UDP-葡糖醛酸供体);EC2.4.1.35(使用UDP- 葡萄糖供体);EC2.4.1.159(使用UDP-鼠李糖供体);EC2.4.1.203(使用UDP- 葡萄糖和/或UDP-木糖供体);EC2.4.1.234(使用UDP-半乳糖供体); EC2.4.1.236(使用UDP-鼠李糖供体)和/或EC2.4.1.294(使用UDP-半乳糖供体) 的那些是特别有用的。
更进一步有用的糖基转移酶是大麻素苷元O-糖基转移酶和/或大麻素糖苷O-糖基转移酶,任选地与SEQ ID NO:1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、101、103、105、107、 109、111、113、115、117、119、121、123、125、127、129、131、133、 135、137、139、141、143、145、147、149、151、153、155、157、159、 161、163、165、167、169、171、173、175、177、179、181、183、185、 187、189、191、193、195、197、199、201、203、205或207中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少 90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O- 糖基转移酶和/或大麻素糖苷O-糖基转移酶。
更进一步有用的糖基转移酶与SEQ ID NO:107、109、111、113、117、 119、121、125、127、129、131、133、135、137、139、141、143、147、 149、151、153、155、157、159、161、163、165、167、169、171、173、 175、177、179、181、183、185、187、189、191、193、195、197、199、 201、203、205、207中任一项包括的大麻素苷元O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
更进一步有用的糖基转移酶是大麻素糖苷O-糖基转移酶,任选地与 SEQ ID NO:115、123或145中任一项包括的大麻素糖O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素糖苷O-糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元O-葡糖基转移酶,任选地与 SEQ IDNO:107、109、111、117、119、121、125、127、129、131、133、 135、137、139、141、143、147、149、151、153、155、157、159、161、 163、165、167、169、171、173、175、177、179、181、183、185、187、189、191、193、195、197、199、201、203、205或207中任一项包括的大麻素苷元O-葡糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-葡糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元O-鼠李糖基转移酶,任选地与SEQ IDNO:107、125、127、147、149、151、157、159、161、177、183、 191、197或207中任一项包括的大麻素苷元O-鼠李糖基转移酶具有至少 70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-鼠李糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元O-木糖基转移酶,任选地与 SEQ IDNO:107、113、125、127、147、149、151、157、159、161、177、 183、191、197或207中任一项包括的大麻素苷元O-木糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-木糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元O-阿拉伯糖基转移酶,任选地与SEQ IDNO:107、125、127、147、149、151、157、159、161、177、 183、191、197或207中任一项包括的大麻素苷元O-阿拉伯糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-阿拉伯糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元O-N-乙酰氨基半乳糖基转移酶,任选地与SEQ ID NO:107、125、127、147、149、151、157、159、161、 177、183、191、197或207中任一项包括的大麻素苷元O-N-乙酰氨基半乳糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-N-乙酰氨基半乳糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元O-N-乙酰氨基葡糖基转移酶,任选地与SEQ ID NO:107、125、127、147、149、151、157、159、161、 177、183、191、197或207中任一项包括的大麻素苷元O-N-乙酰氨基葡糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-N-乙酰氨基葡糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元/糖苷二-O-糖基转移酶,任选地与SEQID NO:107、115、123、125、127、133、135、145、149、151、 157、159、161、165、167、173、175、177、185、191、195或207中任一项包括的大麻素苷元/糖苷二-O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元/糖苷二-O-糖基转移酶。
更进一步有用的糖基转移酶是大麻素苷元/糖苷三-O-糖基转移酶,任选地与SEQID NO:107、115、123、145、157、159、191或207中任一项包括的大麻素苷元/糖苷三-O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元/糖苷三-O-糖基转移酶。
更进一步有用的糖基转移酶是四-O-糖基转移酶,任选地与SEQ ID NO: 207中任一项包括的大麻素苷元/糖苷四-O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的四-O-糖基转移酶。
根据CAZY系统将糖基转移酶分组为不同的家族是技术人员熟知的。在能够糖基化大麻素的糖基转移酶中,属于CAZY体系的酶家族73的糖基转移酶表现特别良好,因此在一种实施方式中,本发明的糖基转移酶是糖基转移酶家族73。特别地,在糖基转移酶家族73中,与SEQ ID NO:107、 157、159、191和/或207中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶表现最佳。
另外表现最佳的糖基转移酶与SEQ ID NO:135、143、147和/或171中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
更进一步有用的糖基转移酶与SEQ ID NO:107、109、111、113、117、 125、127、129、135、137、139、141、147、149、151、153、157、159、 161、177、179、183、191、193、197、201、205或207中任一项包括的糖基化CBD、CBDV和/或CBDA的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性。
更进一步有用的糖基转移酶与SEQ ID NO:107、109、119、125、127、 135、137、147、149、151、157、159、161、165、167、173、175、177、 179、183、185、187、189、191、195、201、205或207中任一项包括的糖基化CBG、CBGV和/或CBGA的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性。
更进一步有用的糖基转移酶与SEQ ID NO:107、111、117、121、125、 127、131、143、149、155、157、159、163、169、171、191、199、201、 203或207中任一项包括的THC糖基化糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
更进一步有用的糖基转移酶与SEQ ID NO:125、127、133、135、149、151、157、159、175、177、181、191、195或207中任一项包括的CBN糖基化糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少 90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
更进一步有用的糖基转移酶与SEQ ID NO:107、125、127、135、149、 151、157、159、175、177、191、201或207中任一项包括的CBC糖基化糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
更进一步有用的糖基转移酶与SEQ ID NO:SEQ ID NO:147、157、107、 159、191、171、135、143中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、例如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
本发明的糖基转移酶与本文所述序列的序列同一性在另外的实施方式中为至少90%,诸如至少95%、诸如至少99%、诸如100%。
在另一种实施方式中,糖基转移酶选自以下中的一种或多种:
a)与SEQ ID NO:1的UGT708G3糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
b)与SEQ ID NO:3的UGT708G2糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
c)与SEQ ID NO:5的UGT708G1糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
d)与SEQ ID NO:7的OsCGT糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性的糖基转移酶;
e)与SEQ ID NO:9的FeUGT708C1糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
f)与SEQ ID NO:11的GmUGT708D1糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
g)与SEQ ID NO:13的ZmUGT708A6糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
h)与SEQ ID NO:15的MiCGT糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
i)与SEQ ID NO:17的GtUF6CGT1糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
j)与SEQ ID NO:19的DcUGT2糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
k)与SEQ ID NO:21的DcUGT4糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
l)与SEQ ID NO:23的DcUGT5糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
m)与SEQ ID NO:25的UGT73B5糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
n)与SEQ ID NO:27的UGT76C5糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
o)与SEQ ID NO:29的UGT73B3糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
p)与SEQ ID NO:31的UGT71E1糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
q)与SEQ ID NO:33的UGT5糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性的糖基转移酶;
r)与SEQ ID NO:35的UGT1A10糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
s)与SEQ ID NO:37的UGT1A9糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
t)与SEQ ID NO:39的UGT2B7糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
更特定地,在一些实施方式中,糖基转移酶选自由以下中的一种或多种组成的组:
a)与SEQ ID NO:31的UGT71E1糖基转移酶具有至少90%(诸如至少 95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
b)与SEQ ID NO:25的UGT73B5糖基转移酶具有至少90%(诸如至少 95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
c)与SEQ ID NO:27的UGT76C5糖基转移酶具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
d)与SEQ ID NO:29的UGT73B3糖基转移酶具有至少90%(诸如至少 95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
e)与SEQ ID NO:33的UGT5糖基转移酶具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
f)与SEQ ID NO:35的UGT1A10糖基转移酶具有至少90%(诸如至少 95%、诸如至少99%、诸如100%)同一性的糖基转移酶;
g)与SEQ ID NO:37的UGT1A9糖基转移酶具有至少90%(诸如至少 95%、诸如至少99%、诸如100%)同一性的糖基转移酶;以及
h)与SEQ ID NO:39的UGT2B7糖基转移酶具有至少90%(诸如至少 95%、诸如至少99%、诸如100%)同一性的糖基转移酶。
在另外的实施方式中,糖基转移酶选自由以下组成的组:
a)与SEQ ID NO:31的UGT71E1糖基转移酶具有至少95%(诸如至少 99%、诸如100%)同一性的糖基转移酶;
b)与SEQ ID NO:25的UGT73B5糖基转移酶具有至少至少95%(诸如至少99%、诸如100%)同一性的糖基转移酶;
c)与SEQ ID NO:27的UGT76C5糖基转移酶具有至少95%(诸如至少 99%、诸如100%)同一性的糖基转移酶;
d)与SEQ ID NO:29的UGT73B3糖基转移酶具有至少95%(诸如至少 99%、诸如100%)同一性的糖基转移酶;
e)与SEQ ID NO:33的UGT5糖基转移酶具有至少95%(诸如至少99%、诸如100%)同一性的糖基转移酶;
f)与SEQ ID NO:35的UGT1A10糖基转移酶具有至少95%(诸如至少99%、诸如100%)同一性的糖基转移酶;
g)与SEQ ID NO:37的UGT1A9糖基转移酶具有至少95%(诸如至少 99%、诸如100%)同一性的糖基转移酶;以及
h)与SEQ ID NO:39的UGT2B7糖基转移酶具有至少95%(诸如至少 99%、诸如100%)同一性的糖基转移酶。
在非限制性实例中,糖基转移酶是:
a)SEQ ID NO:31的UGT71E1糖基转移酶;
b)SEQ ID NO:25的UGT73B5糖基转移酶;
c)SEQ ID NO:27的UGT76C5糖基转移酶;
d)SEQ ID NO:29的UGT73B3糖基转移酶;
e)SEQ ID NO:33的UGT5糖基转移酶;
f)SEQ ID NO:35的UGT1A10糖基转移酶;
g)SEQ ID NO:37的UGT1A9糖基转移酶;或
h)SEQ ID NO:39的UGT2B7糖基转移酶。
本发明的糖基转移酶可以有利地在没有信号肽的情况下表达以避免靶向糖基转移酶进行分泌,并保持其受限于大麻素受体的细胞内糖基化。
进一步有用的糖基转移酶催化糖基基团与大麻素苷元或大麻素糖苷之间形成1,2-、1,3-、1,4-和/或1,6-糖苷键。特别有用的糖基转移酶催化糖基基团与大麻素苷元或大麻素糖苷之间形成1,4-糖苷键和/或1,6-糖苷键。更特别有用的糖基转移酶催化糖基基团与大麻素苷元或大麻素糖苷之间形成 1,4-糖苷键,并且是SEQ ID NO:115中包括的糖基转移酶。替代地,有用的糖基转移酶催化糖基基团与大麻素苷元或大麻素糖苷之间形成1,6-糖苷键,并且是SEQ ID NO:145中包括的糖基转移酶。
基因修饰的细胞包括编码本发明的糖基转移酶的一种或多种异源基因。这些基因可以与编码SEQ ID NO:2、4、6、8、10、12、14、16、18、 20、22、24、26、28、30、32、34、36、38、40、102、104、106、108、 110、112、114、116、118、120、122、124、126、128、130、132、134、 136、138、140、142、144、146、148、150、152、154、156、158、160、 162、164、166、168、170、172、174、176、178、180、182、184、186、188、190、192、194、196、198、200、202、204、206或208中任一项包括的编码糖基转移酶的基因具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。特别有用的基因与SEQ ID NO:148、158、108、160、192、172、137、144中包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少 90%、诸如至少95%、诸如至少99%、诸如100%)同一性。优选地,编码本发明的糖基转移酶的基因与这些选定的序列的序列同一性为至少90%,诸如至少95%、诸如至少99%、诸如100%。更优选地,编码本发明的糖基转移酶的基因与这些选定的序列的序列同一性为至少99%,诸如100%。
在一些实施方式中,编码本发明的糖基转移酶的异源基因选自以下中的一种或多种:
a)与SEQ ID NO:2具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
b)与SEQ ID NO:4具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
c)与SEQ ID NO:6具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
d)与SEQ ID NO:8具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
e)与SEQ ID NO:10具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
f)与SEQ ID NO:12具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
g)与SEQ ID NO:14具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
h)与SEQ ID NO:16具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;以及
i)与SEQ ID NO:18具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
j)与SEQ ID NO:20具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
k)与SEQ ID NO:22具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
l)与SEQ ID NO:24具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
m)与SEQ ID NO:26具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
n)与SEQ ID NO:28具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
o)与SEQ ID NO:30具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
p)与SEQ ID NO:32具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
q)与SEQ ID NO:34具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
r)与SEQ ID NO:36具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
s)与SEQ ID NO:38具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;以及
t)与SEQ ID NO:40具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸。
更具体地,在一些实施方式中,编码糖基转移酶的异源基因选自由以下项中的一种或多种组成的组:
a)与SEQ ID NO:32具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
b)与SEQ ID NO:26具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
c)与SEQ ID NO:28具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
d)与SEQ ID NO:30具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
e)与SEQ ID NO:34具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
f)与SEQ ID NO:36具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
g)与SEQ ID NO:38具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;以及
h)与SEQ ID NO:40具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸。
在进一步的实施方式中,编码糖基转移酶的异源基因选自由以下组成的组:
a)与SEQ ID NO:32具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;
b)与SEQ ID NO:26具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;
c)与SEQ ID NO:28具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;
d)与SEQ ID NO:30具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;
e)与SEQ ID NO:34具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;
f)与SEQ ID NO:36具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;
g)与SEQ ID NO:38具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸;以及
h)与SEQ ID NO:40具有至少95%(诸如至少99%、诸如100%)同一性的多核苷酸。
在非限制性实例中,编码糖基转移酶的异源基因是:
i)SEQ ID NO:32;
j)SEQ ID NO:26;
k)SEQ ID NO:28;
l)SEQ ID NO:30;
m)SEQ ID NO:34;
n)SEQ ID NO:36;
o)SEQ ID NO:38;或
p)SEQ ID NO:40。
大麻素糖苷
本发明包括所有大麻素糖苷,它们是具有上述糖基基团的上述大麻素受体的组合。使用本发明的糖基转移酶,可以产生先前未知的糖基化大麻素,其具有一系列期望的特性,和/或以更有效的方式产生已知的糖基化大麻素。
那些有吸引力的大麻素糖苷具有的水溶解度比相应的未糖基化的大麻素高至少10%。这样的大麻素糖苷包括具有的水溶解度比相应的未糖基化的大麻素高至少10%、至少20%、至少40%、至少60%、至少80%、至少 100%、至少200%和至少500%的大麻素糖苷。可以通过使用本发明的大麻素糖基转移酶制备的一些大麻素糖苷表现出水溶解度增加到最高达相应的未糖基化的大麻素水溶解度的25倍,诸如最高达50倍、诸如最高达100 倍、诸如最高达250倍、诸如最高达500倍、诸如最高达1000倍。对于一些大麻素糖苷,增加的水溶解度可以是相应的未糖基化的大麻素的水溶解度的1000倍以上。增加的水溶解度不仅对发酵生产,而且对向患者给药该产品有巨大的有益影响。
其他有吸引力的大麻素糖苷包括对UV或热降解的抗性比相应的未糖基化的大麻素高至少10%的那些。这样的大麻素糖苷包括对UV或热降解的抗性比相应的未糖基化的大麻素高至少10%、至少20%、至少40%、至少60%、至少80%、至少100%、至少200%和至少500%的大麻素糖苷。又其他有吸引力的大麻素糖苷包括哺乳动物中口服摄取量比相应的未糖基化的大麻素高至少10%的那些,例如当同等地给药于哺乳动物时。这样的大麻素糖苷包括口服摄取量比相应的未糖基化的大麻素高至少20%、至少 40%、至少60%、至少80%、至少100%、至少200%和至少500%的大麻素糖苷。在该背景下,口服摄取量应理解为在胃肠道中吸收至身体血浆中的大麻素糖苷的口服摄入剂量的百分比。又其他有吸引力的大麻素糖苷包括哺乳动物体内的生物半衰期比相应的未糖基化的大麻素高至少10%的那些,例如当同等地给药于哺乳动物时。这种大麻素糖苷包括生物半衰期比相应的未糖基化的大麻素高至少20%、至少40%、至少60%、至少80%、至少100%、至少200%和至少500%的大麻素糖苷。又其他有吸引力的大麻素糖苷包括哺乳动物脑脊液中的峰值浓度的浓度比相应的未糖基化的大麻素高至少10%的那些,例如当同等地给药于哺乳动物时。这种大麻素糖苷包括哺乳动物脑脊液中的峰值浓度的浓度比相应的未糖基化的大麻素高至少20%、至少40%、至少60%、至少80%、至少100%、至少200%和至少 500%的大麻素糖苷。又其他有吸引力的大麻素糖苷包括与相应的未糖基化的大麻素相比药代动力学改善至少10%的那些,例如当同等地给药于哺乳动物时。这样的大麻素糖苷包括与相应的未糖基化的大麻素相比药代动力学改善至少20%、至少40%、至少60%、至少80%、至少100%、至少200%和至少500%的大麻素糖苷,如通过溶解度测定、化学稳定性测定、Caco-2 双向渗透性测定、肝微粒体清除测定和/或血浆稳定性测定所测量的。又其他有吸引力的大麻素糖苷包括与相应的未糖基化的大麻素相比在酸性水溶液中的稳定性提高至少10%的那些,任选地在pH值为0至7(诸如pH值为 0.5至4、诸如pH值为0.5至2、诸如pH值为约1)溶液中。又其他有吸引力的大麻素糖苷包括与相应的未糖基化的大麻素相比在碱性水溶液中稳定性提高至少10%的那些,任选在pH值为7至14(诸如pH值为9至14、诸如pH值为10至13、诸如pH值为约12.5)的溶液中。又其他有吸引力的大麻素糖苷包括与相应的未糖基化的大麻素相比在水溶液中的抗氧化性提高至少10%的那些,任选地在具有至少8mg/L O2(诸如至少20mg/LO2、诸如至少40mg/L O2、诸如至少80mg/L O2)的溶液,诸如诸如用O2饱和的溶液中。又其他有吸引力的大麻素糖苷包括与相应的非糖基化的大麻素相比对基因修饰的宿主细胞的毒性降低至少10%的那些,任选地具有比相应的未糖基化的大麻素低至少10%(诸如低至少25%、诸如低至少75%、诸如低至少100%)的LC50。
在一些实施方式中,大麻素糖苷是C-糖苷或O-糖苷或其组合,特别是选自以下的糖苷的这样的大麻素糖苷:大麻色烯型(CBC)、大麻萜酚型 (CBG)、大麻二酚型(CBD)、四氢大麻酚型(THC)、大麻环酚型(CBL)、大麻艾尔松型(CBE)、大麻酚型(CBN)、脱氢大麻二酚型(CBND)和二羟基大麻酚型大麻素受体。特别有用的大麻素糖苷选自以下的糖苷:大麻二酚(CBD)、大麻二酚酸(CBDA)、次大麻二酚(CBDV)、四氢大麻酚(THC)、四氢大麻酚酸(THCA)、四氢次大麻酚(THCV)、次大麻色烯(CBCV)、大麻萜酚(CBG)、大麻酚(CBN)、11-nor-9-羧基-THC和Δ8-四氢大麻酚。更进一步特别有用的大麻素糖苷选自大麻素-1'-O-β-D-糖苷、大麻素-1'-O-β-D-糖基-3'-O-β-D-糖苷和大麻素-3'-O-β-D-糖苷。更进一步特别有用的大麻素糖苷选自 CBD-1'-O-β-D-糖苷、CBD-1'-O-β-D-糖基-3'-O-β-D-糖苷、CBDV-1’-O-β-D- 糖苷、CBDV-1'-O-β-D-糖基-3'-O-β-D-糖苷、CBG-1'-O-β-D-糖苷、 CBG-1'-O-β-D-糖基-3'-O-β-D-糖苷、THC-1'-O-β-D-糖苷、CBN-1'-O-β-D-糖苷、11-nor-9-羧基-THC-1’-O-β-D-糖苷、CBDA-1-O-β-D-糖苷和 CBC-1’-O-β-D-糖苷。更进一步特别有用的大麻素糖苷选自大麻素糖苷;大麻素葡糖醛酸苷;大麻素木糖苷;大麻素鼠李糖苷;大麻素半乳糖苷;大麻素N-乙酰氨基葡糖苷;大麻素N-乙酰氨基半乳糖苷和大麻素阿拉伯糖苷。又进一步特别有用的大麻素糖苷选自大麻素-1'-O-β-D-葡糖苷;大麻素 -1'-O-β-D-葡糖醛酸苷;大麻素-1'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖苷;大麻素-1'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰氨基葡糖苷;大麻素 -1'-O-β-D-阿拉伯糖苷;大麻素-1'-O-β-D-N-乙酰半乳糖胺;大麻素-1'-O-β-D- 葡糖基-3'-O-β-D-葡糖苷;大麻素-1'-O-β-D-纤维二糖苷;大麻素-1'-O-β-D- 龙胆二糖苷;大麻素-1'-O-β-D-葡糖醛酸基-3'-O-β-D-葡糖醛酸苷;大麻素 -1'-O-β-D-木糖基-3'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖基-3'-O-β-D-鼠李糖苷;大麻素-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N- 乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖基 -3'-O-β-D-阿拉伯糖苷;和大麻素-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺。
产生大麻素受体的起作用的生物合成代谢途径
宿主细胞可以被有利地进一步修饰以包括在从前体产生大麻素受体的途径中产生一种或多种酶的基因。该途径的流程图如图1所示。宿主细胞可以包括从简单营养底物(诸如葡萄糖)产生大麻素受体所需的所有多肽,其从发酵培养基补料。然而,由于底物和前体还可以外源地提供至宿主细胞,并且宿主细胞途径可以包括所选的途径多肽的任何组合,这取决于外源提供的前体和期望由宿主细胞产生的化合物。从单糖到碱性前体乙酰-CoA和丙二酰基-CoA的途径的上游部分是本领域熟知的,例如来自van Rossum等人,2016和Shi等人,2014。此外,从单糖到脂肪酸(诸如己酸)的途径的上游部分也是本领域熟知的,例如来自Gajewski等人,2017或 WO2016156548。在这些基本前体的下游,在一种实施方式中基因修饰的宿主细胞包括起作用的生物合成代谢途径,其包括选自以下的一种或多种多肽:
a)乙酰乙酰-CoA硫解酶(ACT),所述ACT将乙酰-CoA前体转化为乙酰乙酰-CoA;
b)HMG-CoA合酶(HCS),所述HCS将乙酰乙酰-CoA前体转化为 HMG-CoA;
c)HMG-CoA还原酶(HCR),所述HCR将HMG-CoA前体转化为甲羟戊酸;
d)甲羟戊酸激酶(MVK),所述NVK将甲羟戊酸前体转化为甲羟戊酸 -5-磷酸;
e)磷酸甲羟戊酸激酶(PMK),所述PMK将甲羟戊酸-5-磷酸前体转化为甲羟戊酸二磷酸;
f)甲羟戊酸焦磷酸脱羧酶(MPC),所述MPC将甲羟戊酸二磷酸前体转化为异戊烯基二磷酸(IPP);
g)异戊烯基二磷酸/二甲基烯丙基二磷酸异构酶(IPI),所述IPI将IPP 前体转化为二甲基烯丙基二磷酸(DMAPP);
h)香叶基二磷酸合酶(GPPS),所述GPPS将IPP和DMAPP缩合成香叶基二磷酸(GPP);
i)酰基活化酶(AAE),所述AAE将脂肪酸前体转化为脂肪酰基-COA;
j)3,5,7-三氧亚基十二烷酰基-CoA合酶(TKS),所述TKS将脂肪酸-CoA 前体转化为3,5,7-三氧亚基十一烷酰基-CoA;
k)橄榄酸环化酶(OAC),所述OAC将3,5,7-三氧亚基十一烷酰基-CoA 前体转化为divarinolic acid;
l)橄榄酸环化酶(OAC),所述OAC将3,5,7-三氧亚基十二烷酰基-CoA 前体转化为橄榄酸;
m)TKS-OAC融合酶,所述TKS-OAC融合酶将脂肪酸-CoA前体转化为3,5,7-三氧亚基十一烷酰基-CoA、将3,5,7-三氧亚基十一烷酰基-CoA前体转化为divarinolic acid和将3,5,7-三氧亚基十二烷酰基-CoA前体转化为橄榄酸;
n)大麻萜酚酸合酶(CBGAS),所述CBGAS将GPP和橄榄酸缩合为大麻萜酚酸(CBGA);
o)大麻萜酚酸合酶(CBGAS),所述CBGAS将GPP和divarinolic acid 缩合为次大麻萜酚酸(CBGVA);
p)大麻二酚酸合酶(CBDAS),所述CBDAS分别将CBGA酸和/或 CBGVA转化为大麻二酚酸(CBDA)和/或次大麻二酚酸(CBDVA);
q)四氢大麻酚酸合酶(THCAS),所述THCAS分别将CBGA和/或 CBGVA转化为四氢大麻酚酸(THCA)和/或四氢次大麻酚酸(THCVA);
r)大麻色烯酸合酶(CBCAS),所述CBCAS分别将CBGA和/或CBGVA 转化为大麻色烯酸(CBCA)和/或次大麻色烯酸(CBCVA);
s)核苷酸-葡萄糖合酶,所述核苷酸-葡萄糖合酶将蔗糖和核苷酸转化为果糖和核苷酸-葡萄糖;
t)核苷酸-半乳糖4-差向异构酶,所述核苷酸-半乳糖4-差向异构酶将核苷酸-葡萄糖转化为核苷酸-半乳糖;
u)核苷酸-(葡糖醛酸)-脱羧酶,所述核苷酸-(葡糖醛酸)-脱羧酶将核苷酸-葡糖醛酸转化为核苷酸-木糖;
v)核苷酸-4-酮-6-脱氧-葡萄糖3,5-差向异构酶和核苷酸-4-酮-鼠李糖4- 酮-还原酶,它们一起将核苷酸-4-酮-6-脱氧-葡萄糖和NADPH转化为核苷酸-鼠李糖和NADP+;
w)核苷酸-葡萄糖4,6-脱水酶,该核苷酸-葡萄糖4,6-脱水酶将核苷酸- 葡萄糖和NAD+化为核苷酸-4-酮-6-脱氧-葡萄糖和NADH;
x)核苷酸-葡萄糖4,6-脱水酶和核苷酸-4-酮-6-脱氧-葡萄糖3,5-差向异构酶以及核苷酸-4-酮-鼠李糖-4-酮-还原酶,它们一起将核苷酸-葡萄糖和 NAD+以及NADPH转化为核苷酸-鼠李糖+NADH+NADP+;
y)核苷酸-葡萄糖6-脱氢酶,将核苷酸-葡萄糖和2NAD+转化为核苷酸-葡糖醛酸和2NADH;
z)核苷酸-阿拉伯糖4-差向异构酶,将核苷酸-木糖转化为核苷酸-阿拉伯糖;以及
aa)核苷酸-N-乙酰葡糖胺4-差向异构酶,将核苷酸-N-乙酰葡糖胺转化为核苷酸-N-乙酰半乳糖胺。
步骤的核苷酸-葡萄糖合酶也被称为蔗糖合酶,因为它也具有催化可逆反应的能力。
作为途径中可能包括的特定酶的实例,
a)ACT与酿酒酵母中的天然Erg10具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如 100%)同一性;
b)HCS与酿酒酵母中的天然Erg13具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如 100%)同一性;
c)HCS与酿酒酵母中的天然HMG1或HMG2具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
d)MVK与酿酒酵母中的天然Erg12具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如 100%)同一性;
e)PMK与酿酒酵母中的天然Erg8具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
f)MPC与酿酒酵母中的天然MVD1具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如 100%)同一性;
g)IPI与酿酒酵母中的天然IDI1具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
h)GPPS与SEQ ID NO:45或229中包括的GPPS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少 99%、诸如100%)同一性;
i)AAE与SEQ ID NO:47或239中包括的AAE具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少 99%、诸如100%)同一性;
j)TKS与SEQ ID NO:49中包括的TKS具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
k)OAC与SEQ ID NO:51中包括的OAC具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
l)TKS-OAC融合酶与SEQ ID NO:227中包括的TKS-OAC融合酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
m)CBGAS与SEQ ID NO:53、235、237中包括的CBGAS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
n)CBDAS与SEQ ID NO:57或233中包括的CBDAS具有至少 70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
o)THCAS与SEQ ID NO:55或231中包括的THCAS具有至少 70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
p)CBCAS与SEQ ID NO:59中包括的CBCAS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少 99%、诸如100%)同一性;
q)核苷酸-葡萄糖合酶是UDP-葡萄糖合酶并且与SEQ ID NO:209 中包括的UDP-葡萄糖合酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
r)核苷酸-半乳糖4-差向异构酶是UDP-半乳糖4-差向异构酶并且与 SEQ ID NO:211中包括的UDP-半乳糖4-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少 99%、诸如100%)同一性;
s)核苷酸-(葡糖醛酸)-脱羧酶是UDP-葡糖醛酸脱羧酶并且与SEQ ID NO:213中包括的UDP-葡糖醛酸脱羧酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
t)核苷酸-4-酮-6-脱氧-葡萄糖3,5-差向异构酶是UDP-4-酮-6-脱氧- 葡萄糖3,5-差向异构酶并且与SEQ ID NO:215或219中包括的UDP-4- 酮-6-脱氧-葡萄糖3,5-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
u)核苷酸-4-酮-鼠李糖-4-酮还原酶是UDP-4-酮-鼠李糖-4-酮还原酶并且与SEQID NO:215或219中包括的UDP-4-酮-鼠李糖-4-酮还原酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少 95%、诸如至少99%、诸如100%)同一性;
v)核苷酸-葡萄糖4,6脱水酶是UDP-葡萄糖4,6-脱水酶并且与SEQ ID NO:217或219中包括的UDP-葡萄糖4,6-脱水酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少 99%、诸如100%)同一性;
w)核苷酸-葡萄糖6-脱氢酶是UDP-葡萄糖6-脱氢酶并且与SEQ ID NO:221中包括的UDP-葡萄糖6-脱氢酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
x)核苷酸-阿拉伯糖4-差向异构酶是UDP-阿拉伯糖4-差向异构酶并且与SEQ IDNO:223中包括的UDP-阿拉伯糖4-差向异构酶具有至少 70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;以及
y)核苷酸-N-乙酰葡糖胺4-差向异构酶是UDP-N-乙酰葡糖胺4-差向异构酶并且与SEQ ID NO:225中包括的UDP-N-乙酰葡糖胺4-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
SEQ ID NO:232和SEQ ID NO:230两者均是含有液泡定位标签(氨基酸1-24)的N端截短多肽。SEQ ID NO:215包括差向异构酶和还原酶两者,而SEQ ID NO:219包括差向异构酶和还原酶酶(氨基酸1-370)以及脱水酶 (氨基酸371-667)。
更具体地,在另外的实施方式中
a)ACT是酿酒酵母中的天然Erg10;
b)HCS是酿酒酵母中的天然Erg13;
c)HCR是酿酒酵母中的天然HMG1;
d)HCR是酿酒酵母中的天然HMG2;
e)MVK是酿酒酵母中的天然Erg12;
f)PMK是酿酒酵母中的天然Erg8;
g)MPC是酿酒酵母中的天然MVD1;
h)IPI是酿酒酵母中的天然IDI1;
i)GPPS是SEQ ID NO:45或229的GPPS;
j)AAE是SEQ ID NO:47或239的AAE;
k)TKS是SEQ ID NO:49的TKS;
l)OAC是SEQ ID NO:51的OAC;
m)TKS-OAC融合酶是SEQ ID NO 227中包括的TKS-OAC融合酶
n)CBGAS是SEQ ID NO:53、235或237的CBGAS;
o)CBDAS是SEQ ID NO:57或233的CBDAS;
p)THCAS是SEQ ID NO:55或231的THCAS;
q)CBCAS是SEQ ID NO:59的CBCAS;
r)UDP-葡萄糖合酶是SEQ ID NO:209中包括的UDP-葡萄糖合酶;
s)UDP-半乳糖4-差向异构酶是SEQ ID NO:211中包括的UDP-半乳糖4-差向异构酶;
t)UDP-葡糖醛酸脱羧酶是SEQ ID NO:213中包括的UDP-葡糖醛酸脱羧酶;
u)UDP-4-酮-6-脱氧-葡萄糖3,5-差向异构酶是SEQ ID NO:215或 219中包括的UDP-4-酮-6-脱氧-葡萄糖3,5-差向异构酶;
v)UDP-4-酮-鼠李糖-4-酮还原酶是SEQ ID NO:215或219中包括的 UDP-4-酮-鼠李糖-4-酮还原酶;
w)UDP-葡萄糖4,6-脱水酶是SEQ ID NO:217或219中包括的 UDP-葡萄糖4,6-脱水酶;
x)UDP葡萄糖6-脱氢酶是SEQ ID NO:221中包括的UDP-葡萄糖 6-脱氢酶;
y)UDP-阿拉伯糖4-差向异构酶是SEQ ID NO:223中包括的UDP- 阿拉伯糖4-差向异构酶;以及
z)UDP-N-乙酰葡糖胺4-差向异构酶是SEQ ID NO:225中包括的 UDP-N-乙酰葡糖胺4-差向异构酶。
Erg10的序列可见于可公开获得的酵母菌基因组数据库 (www.yeastgenome.org)的SGD ID:SGD:S000005949下;Erg13的序列在 SGD ID:SGD:S000004595下;HMG1的序列在SGD ID:SGD:S000004540 下;HMG2的序列在SGD ID:SGD:S000004442下;Erg12的序列在SGDID: SGD:S000004821下;Erg8的序列在SGD ID:SGD:S000004833下;MVD1 的序列在SGD ID:SGD:S000005326下以及IDI1的序列在SGD ID: SGD:S000006038下。
此外,用于制备大麻素受体的起作用的生物合成代谢途径中包括的多种多肽可以与基因修饰的宿主细胞异源。在更特定的实施方式中,途径多肽中的2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、 19或20种可以与宿主细胞异源。
基因修饰的宿主细胞还可以进一步修饰以优化其大麻素受体的产生。例如,细胞可以被基因修饰以增加起作用的生物合成代谢途径的一种或多种多肽的一种或多种底物或前体或产物的量。这种修饰包括,但不限于合并和表达编码大麻素受体途径和/或编码糖基转移酶的多肽的两个或更多个拷贝,诸如3、4、5或6个拷贝。细胞还可以是基因修饰的宿主细胞,其进一步被基因修饰以对来自起作用的生物合成代谢途径的一种或多种底物、前体、中间体或产物分子表现出增加的耐受性。在又进一步的实施方式中,基因修饰的宿主细胞被修饰以包括促进细胞内形成的大麻素糖苷的分泌的异源转运蛋白多肽。在一些实施方式中,一种或多种天然基因在基因修饰的宿主细胞中被减毒、破坏和/或缺失。例如,在基因修饰的宿主细胞是酿酒酵母菌株的情况下,SGD ID SGD:S000005979的PDR12基因可以被减毒、破坏和/或缺失。
在一些实施方式中,基因修饰的宿主细胞包括公开的多核苷酸构建体或表达载体,见下文。
宿主细胞
基因修饰的宿主细胞可以是任何微生物细胞,诸如真核细胞、原核细胞或古生物细胞。然而,特别有用的宿主细胞是选自由哺乳动物、昆虫、植物或真菌细胞组成的组的真核生物。例如,基因修饰的宿主细胞是大麻和葎草属(Humulus)的植物细胞。在另一种实施方式中,基因修饰的宿主细胞是真菌宿主细胞,其选自以下门:子囊菌门(Ascomycota)、担子菌门 (Basidiomycota)、新丽鞭毛菌门(Neocallimastigomycota)、球囊菌门(Glomeromycota)、芽枝霉门(Blastocladiomycota)、壶菌门(Chytridiomycota)、接合菌门(Zygomycota)、卵菌门(Oomycota)和微孢子菌门(Microsporidia)。更特定地,真菌修饰的宿主细胞可以是选自产子囊孢子酵母(ascosporogenous yeast)(内孢霉目(Endomycetale))、产担孢子酵母(basidiosporogenous yeast)和真菌半知菌酵母(lmperfecti yeast)(芽孢纲(Blastomycete))的酵母细胞。酵母可以选自酵母属(Saccharomyces)、克鲁维酵母菌属(Kluveromyces)、念珠菌属 (Candida)、毕赤酵母属(Pichia)、德巴利氏酵母属(Debaromyces)、汉逊酵母属(Debaromyces)、耶鲁维亚酵母菌(Yarrowia)、接合酵母属(Yarrowia)和裂殖酵母属(Schizosaccharomyces),特别选自由以下组成的种:乳酸克鲁维酵母(Kluyveromyces lactis)、卡尔酵母(Saccharomycescarlsbergensis)、酿酒酵母 (Saccharomyces cerevisiae)、糖化酵母(Saccharomycesdiastaticus)、道格拉斯酵母(Saccharomyces douglasii)、克鲁维酵母(Saccharomyceskluyveri)、诺地酵母(Saccharomyces norbensis)、卵形酵母(Saccharomycesoviformis)、布拉酵母(Saccharomyces boulardii)和解脂耶氏酵母(Yarrowialipolytica)。在另一种实施方式中,基因修饰的宿主细胞是丝状真菌,特别是选自子囊菌门、真菌门和卵菌门的宿主细胞。这样的丝状真菌宿主细胞包括,但不限于选自以下属的那些:支顶孢属(Acremonium)、曲霉属(Aspergillus)、短柄霉属 (Aureobasidium)、烟管菌属(Bjerkandera)、拟蜡菌属(Ceriporiopsis)、金孢子菌属(Chrysosporium)、鬼伞属(Coprinus)、Corio/us、隐球菌属(Cryptococcus)、 Filibasidium、镰刀菌属(Fusarium)、腐质霉属(Humicola)、稻瘟菌属 (Humicola)、毛霉属(Mucor)、毁丝霉属(Myceliophthora)、新美鞭菌属 (Neocallimastix)、链孢霉(Neurospora)、拟青霉属(Paecilomyces)、青霉菌属 (Penicillium)、毛平革菌(Phanerochaete)、白腐菌属(Phlebia)、瘤胃壶菌属 (Piromyces)、侧耳属(Pleurotus)、裂褶菌属(Schizophyllum)、篮状菌属 (Talaromyces)、热子囊菌属(Thermoascus)、梭孢壳属(Thielavia)、弯颈霉属(Tolypocladium)、栓菌属(Trametes)和木霉属(Trichoderma)。在更特定的实施方式中,丝状真菌宿主细胞选自以下的种:泡盛曲霉(Aspergillus awamori)、臭曲霉(Aspergillusfoetidus)、烟曲霉(Aspergillus fumigatus)、日本曲霉 (Aspergillus japonicus)、构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillusoryzae)、烟管菌(Bjerkandera adusta)、干拟蜡菌 (Ceriporiopsis aneirina)、Ceriporiopsis caregiea、Ceriporiopsis gilvescens、潘诺希塔拟蜡菌(Ceriporiopsispannocinta)、环带拟蜡菌(Ceriporiopsis rivulosa)、微红拟蜡菌(Ceriporiopsissubrufa)、虫拟蜡菌(Ceriporiopsis subvermispora)、狭边金孢子菌(Chrysosporiuminops)、嗜角质金孢子菌 (Chrysosporiumkeratinophilum)、卢克诺文思金孢子菌(Chrysosporium lucknowense)、类状金孢子菌(Chrysosporium merdarium)、毡金孢子菌 (Chrysosporium pannicola)、昆士兰金孢子菌(Chrysosporiumqueenslandicum)、热带金孢子菌(Chrysosporium tropicum)、褐薄金孢子菌(Chrysosporium zonatum)、灰盖鬼伞菌(Coprinus cinereus)、毛云芝菌(Coriolushirsutus)、Fusarium bactridioides、禾谷镰孢菌(Fusarium cerealis)、克鲁克威尔镰孢菌(Fusarium crookwellense)、黄色镰孢菌(Fusarium culmorum)、禾谷镰刀菌(Fusariumgraminearum)、禾赤镰孢菌(Fusarium graminum)、异孢镰刀菌(Fusarium heterosporum)、合欢木镰孢菌(Fusarium negundi)、尖孢镰刀菌 (Fusarium oxysporum)、多枝镰孢菌(Fusarium reticulatum)、粉红镰孢菌 (Fusarium roseum)、接骨木镰孢菌(Fusariumsambucinum)、肤色镰孢菌 (Fusarium sarcochroum)、拟分枝镰孢菌(Fusariumsporotrichioides)、硫色镰孢菌(Fusarium sulphureum)、Fusarium torulosum、拟丝孢镰孢菌(Fusarium trichothecioides)、Fusarium venenatum、特异腐质霉(Humicolainsolens)、柔毛腐质霉(Humicola lanuginosa)、米黑毛霉(Mucor miehei)、嗜热毁丝霉(Myceliophthora thermophila)、粉色面包霉菌(Neurospora crassa)、产紫青霉(Penicillium purpurogenum)、黄孢原毛平革菌(Phanerochaete chrysosporium)、射脉齿菌(Phlebia radiata)、刺芹侧耳(Pleurotus eryngii)、土生梭孢壳霉 (Thielaviaterrestris)、长绒毛栓菌(Trametes villosa)、变色栓菌(Trametes versicolor)、哈茨木霉(Trichoderma harzianum)、康宁木霉(Trichoderma koningii)、长枝木霉(Trichodermalongibrachiatum)、里氏木霉(Trichoderma reesei)和绿色木霉(Trichoderma viride)。此外,宿主细胞还可以是三孢布拉霉菌(Blakeslea trispora)。
本发明的基因修饰的宿主细胞还可以是原核细胞,诸如细菌。因此,宿主细胞可以是选自以下属的细菌:埃希氏菌属、乳杆菌属、乳球菌属、棒状菌属、醋杆菌属、不动杆菌属、假单胞菌属或红杆菌属。特别地,宿主细胞可以选自以下种:大肠埃希氏菌(Escherichiacoli)、球形红杆菌 (Rhodobacter sphaeroides)、荚膜红杆菌(Rhodobacter capsulatus)或环红酵母菌(Rhodotorula toruloides)。在一种实施方式中,细菌是大肠埃希氏菌。在另一替代地实施方式中,本发明的宿主细胞是蓝细菌(cyanobacterium)。
本发明的基因修饰的宿主细胞还可以是古生物细胞,诸如藻类。因此,宿主细胞可以选自杜氏盐藻(Dunaliella salina)、雨生红球藻(Haematococcus pluvialis)、小球藻(Chlorella sp.)、裙带菜(Undaria pinnatifida)、马尾藻(Sargassum)、海带(Laminariajaponica)、Scenedesmus almeriensis。
替代地,宿主细胞可以是植物细胞,例如大麻属、葎草属或小孢子藻(Physcomitrella)属的植物细胞。除了植物细胞之外,本发明还提供了分离的植物,例如转基因植物,植物部分包括本发明的大麻素受体途径多肽和糖基转移酶,并产生有效量的本发明的大麻素糖苷。可以从植物或植物部分回收化合物。转基因植物可以是双子叶植物(dicot)或单子叶植物(monocot)。单子叶植物的实例是草类,诸如草地早熟禾(蓝草,早熟禾)、牧草(诸如羊茅、黑麦草)、温带草(例如剪股颖)和谷类,例如小麦、燕麦、黑麦、大麦、水稻、高粱和玉蜀黍(玉米)。双子叶植物的实例是烟草、豆类,例如羽扇豆、马铃薯、甜菜、豌豆、菜豆和大豆,以及十字花科植物(十字花科),诸如花椰菜、油菜籽和密切相关的模式生物拟南芥。植物部分的实例是茎、愈伤组织、叶、根、果实、种子和块茎以及包含这些部分的单独的组织,例如表皮、叶肉、薄壁组织、维管组织、分生组织。特定的植物细胞区室(诸如叶绿体、质外体、线粒体、液泡、过氧化物酶体和细胞质)也被认为是植物部分。此外,任何植物细胞,无论组织来源如何,均被认为是植物部分。同样,为了促进本发明的利用而分离的植物部分(诸如特定组织和细胞)也被认为是植物部分,例如胚、胚乳、糊粉和种皮。此类植物、植物部分和植物细胞的任何后代也包括在本发明的范围内。包括本发明的起作用的途径并产生本发明的化合物的转基因植物或植物细胞可以根据本领域已知的方法构建。简而言之,植物或植物细胞通过以下构建:将一种或多种本发明的表达载体并入至植物宿主基因组或叶绿体基因组中并将所得修饰的植物或植物细胞繁殖成转基因植物或植物细胞。表达载体易于包括本发明的多核苷酸构建体。调节序列(诸如启动子和终止子序列以及任选的信号或转运序列)的选择,例如基于期望表达途径多肽的时间、地点和方式来确定。例如,编码途径酶多肽的基因的表达可以是组成型或诱导型,或者可以是发育、阶段或组织特异性的,并且基因产物可以靶向特定组织或植物部分(诸如种子或叶)。调节序列例如,描述于Tague等人,1988,植物生理学86:506 (Tague et al.,1988,Plant Physiology 86:506)。对于组成型表达,可以使用 358-CaMV、玉米泛素1或水稻肌动蛋白1启动子(Franck等人,1980,细胞21:285-294(Franck et al.,1980,Cell 21:285-294);Christensen等人,1992,植物分子生物学18:675-689(Christensen et al.,1992,Plant Mol.Biol.18: 675-689);Zhang等人,1991,植物细胞3:1155-1165(Zhang et al.,1991,Plant Cell 3:1155-1165))。器官特异性启动子可以是,例如,来自贮藏库组织,诸如种子、马铃薯块茎和果实的启动子(Edwards和Coruzzi,1990,遗传学年鉴24:275-303(Edwards and Coruzzi,1990,Ann.Rev.Genet.24:275-303)),或来自代谢库组织,诸如分生组织(Ito等人,1994,植物分子生物学24: 863-878(Ito et al.,1994,Plant Mol.Biol.24:863-878)),种子特异性启动子,诸如来自水稻的谷蛋白、谷醇溶蛋白、球蛋白或白蛋白启动子(Wu等人, 1998,植物细胞生理学39:885-889(Wu et al.,1998,Plant Cell Physiol.39: 885-889)),来自豆球蛋白B4的蚕豆启动子和来自蚕豆的未知种子蛋白基因 (Conrad等人,1998,植物生理学杂志152:708-711(Conrad et al.,1998,J.Plant Physiol.152:708-711)),来自种子油体蛋白的启动子(Chen等人,1998,植物细胞生理学39:935-941(Chen et al.,1998,Plant CellPhysiol.39:935-941)),来自油菜的贮藏蛋白napA启动子,或本领域已知的任何其他种子特异性启动子,例如,如WO 91/14772所述。此外,启动子可以是叶特异性启动子,诸如来自水稻或番茄的rbcs启动子(Kyozuka等人,1993,植物生理学102: 991-1000(Kyozuka etal.,1993,Plant Physiol.102:991-1000))、小球藻病毒腺嘌呤甲基转移酶基因启动子(Mitra和Higgins,1994,植物分子生物学26: 85-93(Mitra and Higgins,1994,PlantMol.Biol.26:85-93))、来自水稻的aldP 基因启动子(Kagaya等人,1995,分子和一般遗传性248:668-674(Kagaya et al.,1995,Mol.Gen.Genet.248:668-674)),或伤口诱导型启动子,诸如马铃薯 pin2启动子(Xu等人,1993,植物分子生物学22:573-588(Xu et al.,1993,Plant Mol.Biol.22:573-588))。同样,启动子可以由非生物处理(诸如温度、干旱或盐度的改变)诱导,或由外源施加的激活启动子的物质(例如乙醇、雌激素、植物激素(诸如乙烯、脱落酸和赤霉酸)和重金属)诱导。启动子增强子元件也可用于实现在植物中更高的表达。例如,启动子增强子元件可以是位于启动子和编码多肽或结构域的多核苷酸之间的内含子。例如,Xu等人,1993,同上,公开了使用水稻肌动蛋白1基因的第一内含子来增强表达。可选择标记基因和表达构建体的任何其他部分可选自本领域可获得的那些。根据本领域已知的常规技术,包括农杆菌介导的转化、病毒介导的转化、显微注射、粒子轰击、基因枪转化和电穿孔,将多核苷酸构建体或表达载体并入至植物基因组中(Gasser等人,1990,科学244:1293(Gasser et al.,1990, Science 244:1293);Potrykus,1990,生物技术8:535(Potrykus,1990, Bio/Technology 8:535);Shimamoto等人,1989,自然338:274(Shimamoto et al.,1989,Nature 338:274))。根癌土壤杆菌(Agrobacteriumtumefaciens)介导的基因转移是用于产生转基因双子叶植物(综述参见Hooykas和Schilperoort,1992,植物分子生物学19:15-38(Hooykas and Schilperoort,1992, PlantMol.Biol.19:15-38))和用于转化单子叶植物的方法,尽管其他转化方法可以用于这些植物。用于产生转基因单子叶植物的方法是对胚胎愈伤组织或发育中的胚胎进行粒子轰击(涂覆有转化DNA的显微金或钨粒子)(Christou,1992,植物杂志2:275-281(Christou,1992,Plant J.2:275-281); Shimamoto,1994,当代生物技术观点5:158-162(Shimamo,1994, Curr.Opin.Biotechnol.5:158-162);Vasil等人,1992,生物技术10:667-674(Vasil et al.,1992,Bio/Technology 10:667-674))。用于转化单子叶植物的替代方法是基于原生质体转化,如Omirulleh等人,1993,植物分子生物学21: 415-428(Omirulleh etal.,1993,Plant Mo/.Biol.21:415-428)所述。额外的转化方法包括美国专利号6,395,966和7,151,204中所述的那些方法(均通过援引以其全文并入本文)。转化后,根据本领域熟知的方法选择并入至本发明的表达载体或多核苷酸构建体的转化体并使其再生为完整植物。通常,转化程序旨在用于在再生期间或在后代中通过使用,例如,两个单独的T-DNA 构建体的共转化或通过特定重组酶对选择基因进行位点特异性切除来选择性消除选择基因。除了用本发明的多核苷酸构建体直接转化特定植物基因型之外,还可以通过将包括该构建体的植物与缺乏该构建体的第二植物杂交来制备转基因植物。例如,可以通过杂交将编码本发明的糖基转移酶的多核苷酸构建体引入特定植物品种,而无需直接转化该既定品种的植物。因此,本发明不仅涵盖从根据本发明转化的细胞直接再生的植物,还包括这样的植物的后代。如本文所用,后代可以指根据本发明制备的亲本植物的任何代的亲子。这样的后代可以包括本发明的多核苷酸构建体。通过将起始品系与供体植物品系杂交授粉,杂交导致将转基因引入植物品系。此类步骤的非限制性实例描述于美国专利号7,151,204。植物可以通过回交转化过程产生。例如,植物包括被称为回交转化基因型、品系、近交或杂种的植物。遗传标记可以用于辅助本发明的一种或多种转基因从一种遗传背景渗入另一种遗传背景。标记辅助选择相对于传统育种具有优势,因为它可以用于避免由表型变异引起的错误。此外,遗传标记可以提供关于特定杂交的个体后代中优良种质的相对程度的数据。例如,当具有期望性状但具有非农学上期望的遗传背景的植物与优良亲本杂交时,可以使用遗传标记来选择不仅具有感兴趣的性状,而且具有相对较大比例的期望种质的后代。以这种方式,将一种或多种性状渗入特定遗传背景所需的代数最小化。
核苷酸构建体
在另外的方面,本发明提供了一种包括编码本发明的糖基转移酶的多核苷酸序列的多核苷酸构建体,其可操作地连接到与糖基编码多核苷酸异源的一个或多个控制序列。
可以以多种方式操作多核苷酸以实现多肽表达。根据表达载体,在将多核苷酸插入表达载体之前对其进行操作可能是期望的或必要的。利用重组DNA方法修饰多核苷酸的技术是本领域熟知的。
控制序列可以是启动子,其是被宿主细胞识别以表达多核苷酸的多核苷酸。启动子包含介导多肽表达的转录控制序列。启动子可以是在宿主细胞中显示转录活性的任何多核苷酸,包括突变、截短和杂合启动子,并且可以从编码与宿主细胞同源或异源的细胞外或细胞内多肽的基因获得。启动子可以是诱导型启动子。
用于指导本发明的多核苷酸构建体在丝状真菌宿主细胞中转录的合适启动子的实例是获自以下的基因的启动子:构巢曲霉乙酰胺酶、黑曲霉中性α-淀粉酶、黑曲霉酸稳定性α-淀粉酶、黑曲霉或泡盛曲霉葡糖淀粉酶 (glaA)、曲霉属gpdA启动子、米曲霉TAKA淀粉酶、米曲霉碱性蛋白酶、米曲霉磷酸丙糖异构酶、黑曲霉或泡盛曲霉木聚糖内切酶(xlnA)或β-木糖苷酶(xlnD)、尖孢镰孢菌胰蛋白酶样蛋白酶(WO 96/00787)、Fusarium venenatum淀粉葡糖苷酶(WO2000/56900)、Fusarium venenatum Dania(WO 00/56900)、Fusariumvenenatum Quinn(WO 00/56900)、米黑根毛霉脂肪酶、米黑根毛霉天冬氨酸蛋白酶、里氏木霉(Trichoderma reesei)β-葡糖苷酶、里氏木霉纤维二糖水解酶I、里氏木霉纤维二糖水解酶II、里氏木霉内切葡聚糖酶I、里氏木霉内切葡聚糖酶Ⅱ、里氏木霉内切葡聚糖酶ⅡI、里氏木霉内切葡聚糖酶IV、里氏木霉内切葡聚糖酶V、里氏木霉木聚糖酶I、里氏木霉木聚糖酶II、里氏木霉β-木糖苷酶,以及NA2-tpi启动子及其突变、截短和杂合启动子。NA2-tpi启动子是来自曲霉属中性α-淀粉酶基因的修饰启动子,其中未翻译的前导序列已被来自曲霉磷酸丙糖异构酶基因的未翻译的前导序列替换。此类启动子的实例包括来自黑曲霉中性α-淀粉酶基因的修饰启动子,其中未翻译的前导序列已被来自构巢曲霉或米曲霉磷酸丙糖异构酶基因的未翻译的前导序列替换。启动子的其他实例是W02006/092396、 W02005/100573和W02008/098933中描述的启动子,其通过援引并入本文。
用于指导本发明的多核苷酸构建体在酵母宿主中转录的合适启动子的实例包括甘油醛-3-磷酸脱氢酶启动子,PgpdA或获自酿酒酵母烯醇化酶 (EN0-1)、酿酒酵母半乳糖激酶(GAL1)、酿酒酵母乙醇脱氢酶/甘油醛-3-磷酸脱氢酶(ADH1,ADH2/GAP)、酿酒酵母磷酸丙糖异构酶(TPI)、酿酒酵母金属硫蛋白(CUP1)和酿酒酵母3-磷酸甘油酸激酶的基因的启动子。用于酵母宿主细胞的其他有用的启动子描述于Romanos等人,1992,酵母8: 423-488(Romanos et al.,1992,Yeast 8:423-488)。选择用于在酵母中表达的合适启动子是本领域技术人员所熟知且很好理解的。
控制序列还可以是转录终止子,其被宿主细胞识别以终止转录。终止子可操作地连接至编码多肽的多核苷酸的3'-末端。可以使用在宿主细胞中是功能性的任何终止子。
对丝状真菌宿主细胞有用的终止子获自以下的基因:构巢曲霉邻氨基苯甲酸合酶、黑曲霉葡糖淀粉酶、黑曲霉α-葡糖苷酶、米曲霉TAKA淀粉酶和尖孢镰孢菌胰蛋白酶样蛋白酶。
酵母宿主细胞的有用终止子是获自酿酒酵母烯醇化酶、酿酒酵母细胞色素C(CYC1)和酿酒酵母甘油醛-3-磷酸脱氢酶的基因。酵母宿主细胞的其他有用终止子描述于Romanos等人,1992,同上。
控制序列还可以是启动子下游和基因编码序列上游的mRNA稳定子区,其增加基因的表达。
控制序列还可以是前导序列,即对宿主细胞翻译重要的mRNA的非翻译区。前导序列可操作地连接至编码多肽的多核苷酸的5'-末端。可以使用在宿主细胞中具有功能性的任何前导序列。
丝状真菌宿主细胞的优选前导序列获自米曲霉TAKA淀粉酶和构巢曲霉磷酸丙糖异构酶的基因。
酵母宿主细胞的合适前导序列获自酿酒酵母烯醇化酶(EN0-1)、酿酒酵母3-磷酸甘油酸激酶、酿酒酵母α-因子和酿酒酵母醇脱氢酶/甘油醛-3-磷酸脱氢酶((ADH2/GAP)的基因。
控制序列还可以是聚腺苷酸化序列;与多核苷酸的3'-末端可操作连接的序列,当转录时,被宿主细胞识别为向转录的mRNA添加多聚腺苷残基的信号。可以使用在宿主细胞中具有功能性的任何聚腺苷酸化序列。
用于丝状真菌宿主细胞的有用聚腺苷酸化序列获自构巢曲霉邻氨基苯甲酸合酶、黑曲霉葡糖淀粉酶、黑曲霉α-葡糖苷酶、米曲霉TAKA淀粉酶和尖孢镰孢菌胰蛋白酶样蛋白酶的基因。
用于酵母宿主细胞的有用聚腺苷酸化序列描述于Guo和Sherman, 1995,分子细胞生物学15:5983-5990(Guo and Sherman,1995,Mol.Cellular Biol.15:5983-5990)。
还可能需要添加调节序列,以相对于宿主细胞的生长调节多肽的表达。调节系统的实例是那些响应化学或物理刺激(包括调节化合物的存在)而导致基因表达开启或关闭的系统。
在丝状真菌中,可以使用黑曲霉葡糖淀粉酶启动子、米曲霉TAKA α- 淀粉酶启动子和米曲霉葡糖淀粉酶启动子。
在酵母中,可以使用ADH2系统或GAL1系统。调节序列的其他实例是那些允许基因扩增的序列。在真核系统中,这些调节序列包括在存在甲氨蝶呤的情况下扩增的二氢叶酸还原酶基因,以及在存在重金属的情况下扩增的金属硫蛋白基因。
在一种实施方式中,编码糖基转移酶的多核苷酸选自:
a)与SEQ ID NO:2具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
b)与SEQ ID NO:4具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
c)与SEQ ID NO:6具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
d)与SEQ ID NO:8具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
e)与SEQ ID NO:10具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
f)与SEQ ID NO:12具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
g)与SEQ ID NO:14具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
h)与SEQ ID NO:16具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;以及
i)与SEQ ID NO:18具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
j)与SEQ ID NO:20具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
k)与SEQ ID NO:22具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
l)与SEQ ID NO:24具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
m)与SEQ ID NO:26具有至少70%(诸如至少75%、诸如至少 80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
n)与SEQ ID NO:28具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
o)与SEQ ID NO:30具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
p)与SEQ ID NO:32具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;以及
q)与SEQ ID NO:34具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的多核苷酸;
在另一种实施方式中,本发明的多核苷酸构建体中编码糖基转移酶的多核苷酸与SEQ ID NO:2、4、6、8、10、12、14、16、18、20、22、24、 26、28、30、32、34、36、38、40、102、104、106、108、110、112、114、 116、118、120、122、124、126、128、130、132、134、136、138、140、142、144、146、148、150、152、154、156、158、160、162、164、166、 168、170、172、174、176、178、180、182、184、186、188、190、192、 194、196、198、200、202、204、206或208中任一项包括编码糖基转移酶的基因具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
表达载体
在另外的方面,本发明提供了一种包括本发明的多核苷酸构建体的表达载体。除了本发明的多核苷酸构建体之外,各种核苷酸序列可以连接在一起以产生重组表达载体,其可以包括一个或多个方便的限制性位点以允许在这些位点插入或取代编码相关多肽的多核苷酸序列。重组表达载体可以是能够方便地进行重组DNA程序并且可以引起编码相关多肽的多核苷酸的表达的任何载体(例如,质粒或病毒)。载体的选择通常取决于载体与待引入载体的宿主细胞的相容性。载体可以是线性或闭环质粒。载体可以是自主复制载体,即作为染色体外实体存在的载体,其复制不依赖于染色体复制,例如质粒、染色体外元件、微型染色体或人工染色体。载体可以包括用于确保自我复制的任何方式。替代地,当被引入宿主细胞时,载体可以整合至基因组中并与其整合的一种或多种染色体一起复制。此外,可以使用单个载体或质粒或者两个或更多个载体或质粒,它们一起包含待引入宿主细胞基因组的总DNA,或转座子。载体可以包含一种或多种可选择标记,其允许容易地选择转化的、转染的、转导的等细胞。可选择标记是产物提供杀生物剂或病毒抗性、重金属抗性、营养缺陷型的原养型等的基因。
丝状真菌宿主细胞的有用可选择标记包括amdS(乙酰胺酶)、argB(鸟氨酸氨基甲酰基转移酶)、bar(膦丝菌素乙酰转移酶)、hph(潮霉素磷酸转移酶)、 niaD(硝酸还原酶)、pyrG(乳清苷-5'-磷酸脱羧酶)、sC(硫酸腺苷酸转移酶)和 trpC(邻氨基苯甲酸合酶),以及其等同物。构巢曲霉(Aspergillus nidulans) 或米曲霉(Aspergillus oryzae)amdS和pyrG基因以及吸水链霉菌 (Streptomyces hygroscopicus)bar基因在曲霉细胞中特别有用。
酵母宿主细胞的有用的可选择标记包括,但不限于ADE2、HIS3、LEU2、 LYS2、MET3、TRP1和URA3。
载体优选包含允许载体整合至宿主细胞基因组中或允许载体在细胞中非依赖于基因组自主复制的元件。为了整合至宿主细胞基因组中,载体可以依靠编码多肽的多核苷酸或载体的任何其他元件通过同源或非同源重组整合至基因组中。替代地,载体可以包含额外的多核苷酸,用于在一种或多种染色体中的一个或多个精确位置通过同源重组指导整合至宿主细胞的基因组中。为了增加在精确位置整合的可能性,整合元件应该包含足够数量的核酸,诸如35至10,000个碱基对,例如100至10,000个碱基对,例如400至10,000个碱基对,以及诸如800至10,000个碱基对,其与相应目标序列具有高度序列同一性,以提高同源重组的概率。整合元件可以是与宿主细胞基因组中的靶序列同源的任何序列。此外,整合元件可以是非编码或编码多核苷酸。另一方面,载体可以通过非同源重组整合至宿主细胞的基因组中。
复制起点可以是在细胞中具有功能性的介导自主复制的任何质粒复制子。术语“复制起点”或“质粒复制子”是指能使质粒或载体在体内复制的多核苷酸。
用于丝状真菌细胞的有用复制起点包括AMA 1和ANS1(Gems等人, 1991,基因98:61-67(Gems et al.,1991,Gene 98:61-67);Cullen等人,1987,核酸研究15:9163-9175(Cullen et al.,1987,Nucleic Acids Res.15: 9163-9175);WO 00/24883)。AMA 1基因的分离和包括该基因的质粒或载体的构建可以使用WO 00/24883中公开的方法完成。
酵母宿主细胞的有用复制起点是2微米复制起点、ARS1、ARS4、ARS1 和CEN3的组合以及ARS4和CEN6的组合。
可以将编码糖基转移酶或本发明的其他途径多肽的多核苷酸的多于一个拷贝插入宿主细胞中以增加多肽的产生。拷贝数增加可以通过将酶编码序列的一个或多个额外的拷贝整合至宿主细胞基因组中或通过包括可扩增的可选择标记基因与多核苷酸来获得,使得可以通过在合适的可选择剂存在下培养细胞来选择包含可选择标记基因的扩增拷贝的细胞以及由此多核苷酸的额外的拷贝。用于连接上述元件以构建本发明的重组表达载体的程序是本领域技术人员熟知的(参见,例如Sambrook等人,1989,同上)。
细胞培养物
在另外的方面,本发明提供了一种包括本发明的基因修饰的宿主细胞和生长培养基的细胞培养物。用于宿主细胞(诸如植物细胞系、丝状真菌和/ 或酵母)的合适的生长培养基是本领域已知的。
产生本发明的化合物的方法。
在另外的方面,本发明提供了一种用于产生大麻素糖苷的方法,包括:
a)在允许基因修饰的宿主细胞产生大麻素糖苷的条件下培养本发明要求的细胞培养物;以及
b)任选地回收和/或分离所述大麻素糖苷。
可以使用本领域已知的方法在适合于产生本发明化合物和/或增殖细胞计数的营养培养基中培养细胞培养物。例如,培养物可以通过摇瓶培养或者小规模或大规模发酵(包括连续、分批、补料分批或固态发酵),在实验室或工业发酵罐中在允许该途径起作用以产生本发明的化合物的合适的培养基和条件下培养并且任选地被回收和/或分离。
使用本领域已知的程序,在包含碳源和氮源以及无机盐的合适营养培养基中进行培养。合适的培养基可商购获得,或可以根据公开的组合物(例如,在美国典型培养物保藏中心的目录中)制备。合适培养基的选择可以基于宿主细胞的选择和/或基于宿主细胞的调节要求。这样的培养基是本领域已知的。如有需要,培养基可以包含有利于转化的表达宿主而不是其他潜在污染微生物的额外的组分。因此,在实施方式中,合适的营养培养基包括碳源(例如葡萄糖、麦芽糖、糖蜜、淀粉、纤维素、木聚糖、果胶、木质纤维素分解生物质水解物等)、氮源(例如硫酸铵、硝酸铵、氯化铵等)、有机氮源(例如酵母提取物、麦芽提取物、蛋白胨等)和无机营养源(例如磷酸盐、镁、钾、锌、铁等)。
宿主细胞的培养可在约0.5天至约30天的时间段内进行。培养过程可以是分批过程、连续或分批补料过程,适合在0-100℃或0-80℃范围内(例如约0℃至约50℃)的温度下和/或在例如约2至约10的pH值下进行。酵母和丝状真菌的优选发酵条件是约25℃至约55℃范围内的温度和约3至约9的pH值。通常基于宿主细胞的选择来选择合适的条件。因此,在实施方式中,本发明的方法进一步包括选自以下的一种或多种要素:
a)在营养培养基中培养细胞培养物;
b)在需氧或厌氧条件下培养细胞培养物
c)在搅拌下培养细胞培养物;
d)在25至50℃的温度下培养细胞培养物;
e)在3-9之间的pH下培养细胞培养物;
c)将细胞培养物培养10小时至30天;以及
d)在分批补料、重复分批补料或半连续条件下培养细胞
e)在有机溶剂存在下培养细胞培养物以提高大麻素苷元的溶解度。
此外,在一种实施方式中,用于产生大麻素糖苷的方法包括大麻素受体和/或大麻素糖苷的非酶促脱羧步骤。脱羧可以通过热处理、UV处理或碱度处理或其组合来实现。
该方法可以进一步包括将一种或多种外源大麻素受体和/或核苷酸-糖苷补料至细胞培养物。
可以使用本领域已知的方法回收和/或分离本发明的大麻素糖苷。例如,可以通过常规程序(包括但不限于收集、离心、过滤、提取、喷雾干燥、蒸发或沉淀)从营养培养基中回收大麻素糖苷。大麻素糖苷可以通过本领域已知的多种程序分离,包括但不限于色谱法(例如,离子交换、亲和、疏水、色谱聚焦和尺寸排阻)、电泳程序(例如,制备式等电聚焦)、差异溶解度(例如,硫酸铵沉淀)、SDS-PAGE或提取(参见,例如,蛋白纯化,Janson和 Ryden,编辑,VCH出版社,纽约,1989(Protein Purification,Janson and Ryden, editors,VCHPublishers,New York,1989))。在特定的实施方式中,本发明方法的回收和/或分离步骤包括将宿主细胞或细胞培养物的液相与宿主细胞或细胞培养物的固相分离以获得包括本发明的大麻素糖苷的上清液,其通过选自以下项的一个或多个步骤:
a)分解基因修饰的宿主细胞以将细胞内的大麻素糖苷释放到上清液中;
b)使上清液与一种或多种吸附树脂接触以获得产生的大麻素糖苷的至少一部分;
c)使上清液与一个或多个离子交换或反相色谱柱接触以获得大麻素糖苷的至少一部分;以及
d)结晶或提取大麻素糖苷;以及
e)蒸发液相溶剂以浓缩或沉淀大麻素糖苷;
从而回收和/或分离大麻素糖苷。
本发明方法的大麻素糖苷在宿主细胞中的产率优选比通过使用来自甜叶菊的糖基转移酶UGT76G1的产率高至少10%,诸如至少50%、诸如至少100%、诸如至少150%、诸如至少200%。
并非产生本发明的大麻素受体的途径的所有转化步骤均需要在宿主细胞的体内发生,因此在特定实施方式中,这些步骤中的一个或多个步骤在体外进行。因此,在实施方式中,本发明的方法包括至少一个在体外进行的大麻素受体途径步骤。
在一种实施方式中,产生大麻素糖苷的方法包括以下步骤:将大麻素糖苷加工成药用大麻素制剂,其包括向包括非植物细胞的本发明的细胞培养物补料在生长培养基中的起始材料;从细胞培养物中产生药用大麻素化合物以产生包含细胞培养物、生长培养基和药用大麻素化合物的混合物;对药物大麻素化合物进行处理,其中处理包括:使用选自由沉降、过滤和离心组成的组中的至少一种方法分离基因修饰的细胞;以及产生包括药用大麻素的药用大麻素制剂,其中混合物不含可检测量的植物杂质,该植物杂质选自由以下组成的组:多糖、木质素、色素、类黄酮、菲类 (phenanthreoids)、胶乳、树胶、树脂、蜡、杀虫剂、杀真菌剂、除草剂和花粉。
在单独的方面,本发明还提供了一种用于产生大麻素糖苷的方法,包括在允许糖基转移酶将核苷酸糖苷的糖基部分转移至大麻素的条件下,使大麻素受体与本发明的一种或多种大麻素糖基转移酶和本发明的一种或多种核苷酸糖苷接触。特别地,该方面的方法可以在本发明的基因修饰的细胞中在体外进行以及体内进行。
2.产生大麻素糖苷的方法可以进一步包括使大麻素糖苷经受一个或多个去糖基化步骤。去糖基化可以通过将大麻素糖苷与一种或多种酶孵育来实现,该酶选自葡糖苷酶、果胶酶、阿拉伯糖酶、纤维素酶、葡聚糖酶、半纤维素酶和木聚糖酶。特别有用的去糖基化酶包括β-葡糖苷酶、β-β葡糖苷酶、果胶裂解酶、果胶酶(pectozyme)和多半乳糖醛酸酶。去糖基化步骤尤其可以在体外进行。
发酵液
在另外的方面,本发明提供了一种发酵液,该发酵液包括本发明的细胞培养物中包括的大麻素糖苷。优选地,至少50%(诸如至少75%、诸如至少95%、诸如至少99%)的经基因修饰的宿主细胞被分解,并且优选地,至少50%(诸如至少75%、诸如至少95%、诸如至少99%)的固体细胞材料已从液体中分离。在实施方式中,发酵液进一步包括选自以下的一种或多种化合物:
a)产生大麻素糖苷的起作用的生物合成代谢途径的前体或产物;
b)包括微量金属、维生素、盐、酵母氮源基础、YNB和/或氨基酸的补充营养物;以及
其中大麻素糖苷的浓度为至少1mg/l发酵液。优选地,发酵液中的大麻素浓度为至少5mg/L,诸如至少10mg/L、诸如至少20mg/l、诸如至少 50mg/L、诸如至少100mg/L、诸如至少500mg/L、诸如至少1000mg/L、诸如至少5000mg/L、诸如至少10000mg/L、诸如至少50000mg/L。
化合物和组合物
已经发现本发明的糖基转移酶可以产生新的有用的大麻素糖苷。因此,在一方面,本发明提供了一种大麻素糖苷,包括与选自以下的糖共价连接的大麻素苷元或大麻素糖苷:木糖;鼠李糖;半乳糖;N-乙酰葡糖胺;N- 乙酰半乳糖胺;和阿拉伯糖。
进一步地,这些大麻素糖苷可以选自CBD-1'-O-β-D-木糖基-3'-O-β-D- 木糖苷;CBD-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷;CBD-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBD-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;CBD-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;CBD-1'-O-β-D-N- 乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺;CBDV-1'-O-β-D-木糖基-3'-O-β-D- 木糖苷;CBDV-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷;CBDV-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBDV-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;CBDV-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷; CBDV-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺;CBG-1'-O-β-D- 木糖基-3'-O-β-D-木糖苷CBG-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷; CBG-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBG-1'-O-β-D-N-乙酰葡糖胺 -3'-O-β-D-N-乙酰氨基葡糖苷;CBG-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;CBG-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺; THC-1'-O-β-D-木糖苷;THC-1'-O-α-L-鼠李糖苷;THC-1'-O-β-D-半乳糖苷; THC-1'-O-β-D-N-乙酰氨基葡糖苷;THC-1'-O-β-D-阿拉伯糖苷; THC-1'-O-β-D-N-乙酰氨基半乳糖苷;CBN-1'-O-β-D-木糖苷;CBN-1'-O-α-L- 鼠李糖苷;CBN-1'-O-β-D-半乳糖苷;CBN-1'-O-β-D-N-乙酰氨基葡糖苷; CBN-1'-O-β-D-阿拉伯糖苷;CBN-1'-O-β-D-N-乙酰氨基半乳糖苷;CBDA-1'-O-β-D-木糖苷;CBDA-1'-O-α-L-鼠李糖苷;CBDA-1'-O-β-D-半乳糖苷;CBDA-1'-O-β-D-N-乙酰氨基葡糖苷;CBDA-1'-O-β-D-阿拉伯糖苷; CBDA-1'-O-β-D-N-乙酰氨基半乳糖苷;CBC-1'-O-β-D-木糖苷; CBC-1'-O-α-L-鼠李糖苷;CBC-1'-O-β-D-半乳糖苷;CBC-1'-O-β-D-N-乙酰氨基葡糖苷;CBC-1'-O-β-D-阿拉伯糖苷;和CBC-1'-O-β-D-N-乙酰氨基半乳糖苷。先前未公开的特别令人感兴趣的大麻素糖苷是通过1,4或1,6-糖苷键与糖基部分共价连接的大麻素苷元或大麻素糖苷。更进一步地,大麻素糖苷可以是CBD-1'-O-β-D-龙胆二糖苷或CBD-1'-O-β-D-纤维二糖苷。
新的大麻素糖苷分子可以分为以下几组,连同催化糖基化的本发明的糖基转移酶的实例。
Figure BDA0003490979370000581
更特定地,催化糖基化的本发明的新的大麻素糖苷分子和糖基转移酶的实例包括:
Figure BDA0003490979370000591
Figure BDA0003490979370000601
Figure BDA0003490979370000611
Figure BDA0003490979370000621
Figure BDA0003490979370000631
Figure BDA0003490979370000641
Figure BDA0003490979370000651
Figure BDA0003490979370000661
Figure BDA0003490979370000671
Figure BDA0003490979370000681
Figure BDA0003490979370000691
Figure BDA0003490979370000701
Figure BDA0003490979370000711
Figure BDA0003490979370000721
Figure BDA0003490979370000731
Figure BDA0003490979370000741
Figure BDA0003490979370000751
Figure BDA0003490979370000761
在另外的方面,本发明提供了一种组合物,该组合物包括本发明的发酵液以及一种或多种剂、添加剂和/或赋形剂。剂、添加剂和/或赋形剂包括制剂添加剂、稳定剂和填充剂。
可以使用本领域已知的方法将本发明的组合物配制成干燥固体形式。此外,组合物可以是干燥形式,诸如使用本领域已知的方法制备的喷雾干燥、喷雾冷却、冻干、速冻、颗粒、微粒、胶囊或微胶囊形式。
还可以使用本领域已知的方法将本发明的组合物配制成液体稳定形式。此外,组合物可以是液体形式,诸如包括一种或多种稳定剂(诸如糖和/ 或多元醇(例如糖醇))和/或有机酸(例如乳酸)的稳定液体。
在一种特定的实施方式中,将组合物精制成适合人或动物摄取的饮料,并且与未糖基化的大麻素相比,大麻素糖苷具有的水溶解度增加。在另一种特定的实施方式中,将组合物精制成适合人或动物摄取的固体食品,并且其中与未糖基化的大麻素相比,大麻素糖苷具有的水溶解度增加。
药物制剂
在另外的方面,本发明提供了一种用于制备药物制剂的方法,所述方法包括将本发明的组合物与一种或多种药物级赋形剂、添加剂和/或佐剂混合。在另外的方面,本发明提供了一种用于制备药物制剂的方法,包括将本发明的新型大麻素糖苷或本发明的组合物与一种或多种药物级赋形剂、添加剂和/或佐剂混合。大麻素糖苷通常用作前药,其中糖基基团在体内裂解,留下大麻素作为活性药物化合物。
药物制剂可以是粉末、片剂、胶囊、硬咀嚼剂和/或软锭剂或口香糖的形式。药物制剂替代地可以是液体药物溶液的形式。
本发明还提供了一种可获自本发明的用于制备药物制剂的方法的药物制剂。在实施方式中,药物制剂可以用作预防、治疗、缓解和/或减轻哺乳动物中疾病的药物或前药。这样的疾病包括但不限于NASH、癫痫、呕吐、恶心、癌症、多发性硬化症、痉挛、慢性疼痛、厌食症、食欲不振、帕金森病、德拉韦综合征(婴儿严重肌阵挛癫痫)、伦诺克斯-加斯托综合征、物质(药物)滥用、糖尿病、癫痫发作、恐慌症、社交焦虑症(SAD)、广泛性焦虑症(GAD)、焦虑症、广场恐惧症、婴儿痉挛症(韦斯特综合征)、银屑病、疱疹后神经痛、运动神经元疾病、肌萎缩侧索硬化、图雷特综合征、抽动障碍、大脑性瘫痪、移植物抗宿主病(GVHD)、克罗恩病(区域性肠炎)、炎症性肠病、脆性X综合征、双相情感障碍(躁狂抑郁症)、骨关节炎、亨廷顿病、精神分裂症、自闭症、不安腿综合征、人类免疫缺陷病毒(HIV)感染 (AIDS)、高血压、肝纤维化、肝损伤、普拉德-威利综合征(PWS)、创伤后应激障碍(PTSD)、脂肪肝、青光眼、炎症性疾病、艰难梭菌感染、结直肠肿瘤、炎症性肠病、肠病、肠易激综合征、溃疡性结肠炎、认知障碍、脑缺氧、纤维化、睡眠呼吸暂停和运动神经元病。其他医学病症包括减轻其他药物的副作用,包括化疗引起的恶心、痉挛、神经性疼痛、头晕、镇静、精神错乱、分离(dissociation)和“情绪高涨”。哺乳动物优选为人、家畜和/ 或宠物。
糖基化大麻素可以作为前药,因为在给药后糖分子可以在身体的不同位置被例如在肝脏、小肠、脾脏和/或肾脏中发现的胞质葡糖苷酶从大麻素受体上切割下来。微生物葡糖苷酶还可以从大麻素受体上切割糖分子,并且这样的微生物可以在例如胃肠道(肠道微生物组)和人唾液(唾液微生物组) 中发现。当糖苷或糖附连至大麻素受体时,该糖苷可能具有生物学惰性,而在从大麻素受体中去除糖时,它可以重新获得其生物活性和治疗效果。
使用方法
在最后一方面,本发明提供了使用本公开的药物制剂治疗哺乳动物中疾病的方法,包括向哺乳动物给药治疗有效量的药物制剂。这样的疾病包括但不限于NASH、癫痫、呕吐、恶心、癌症、多发性硬化症、痉挛、慢性疼痛、厌食症、食欲不振、帕金森病、德拉韦综合征(婴儿严重肌阵挛癫痫)、伦诺克斯-加斯托综合征、物质(药物)滥用、糖尿病、癫痫发作、恐慌症、社交焦虑症(SAD)、广泛性焦虑症(GAD)、焦虑症、广场恐惧症、婴儿痉挛症(韦斯特综合征)、银屑病、疱疹后神经痛、运动神经元疾病、肌萎缩侧索硬化、图雷特综合征、抽动障碍、大脑性瘫痪、移植物抗宿主病(GVHD)、克罗恩病(区域性肠炎)、炎症性肠病、脆性X综合征、双相情感障碍(躁狂抑郁症)、骨关节炎、亨廷顿病、精神分裂症、自闭症、不安腿综合征、人类免疫缺陷病毒(HIV)感染(AIDS)、高血压、肝纤维化、肝损伤、普拉德- 威利综合征(PWS)、创伤后应激障碍(PTSD)、脂肪肝、青光眼、炎症性疾病、艰难梭菌感染、结直肠肿瘤、炎症性肠病、肠病、肠易激综合征、溃疡性结肠炎、认知障碍、脑缺氧、纤维化、睡眠呼吸暂停和运动神经元病。其他医学病症包括减轻其他药物的副作用,包括化疗引起的恶心、痉挛、神经性疼痛、头晕、镇静、精神错乱、分离和“情绪高涨”。
序列
本申请含有在PatentIn 3.5.1版中制成的序列表,其还以ST25格式以电子方式提交,其通过援引以其全文并入本文。
在本公开的全文中,可以使用基因、引物和/或酶的简称或缩写,此类简称与序列标识符相关联,如下所示:
Figure BDA0003490979370000781
Figure BDA0003490979370000791
Figure BDA0003490979370000801
Figure BDA0003490979370000811
Figure BDA0003490979370000821
Figure BDA0003490979370000831
Figure BDA0003490979370000841
Figure BDA0003490979370000851
Figure BDA0003490979370000861
Figure BDA0003490979370000871
Figure BDA0003490979370000881
Figure BDA0003490979370000882
Figure BDA0003490979370000891
Figure BDA0003490979370000901
Figure BDA0003490979370000911
Figure BDA0003490979370000921
Figure BDA0003490979370000931
本发明的列举方面和实施方式
本发明进一步提供了以下实施方式和条目:
1.一种基因修饰以在细胞内产生大麻素糖苷的微生物宿主细胞,所述细胞表达编码至少一种糖基转移酶的异源基因,该糖基转移酶能够使大麻素受体与糖基供体发生细胞内糖基化,从而产生大麻素糖苷。
2.根据条目1所述的基因修饰的宿主细胞,其中大麻素受体是异戊二烯基供体和异戊二烯基受体的缩合产物或其衍生物。
3.根据条目1或2所述的基因修饰的宿主细胞,其中大麻素受体是大麻素苷元或大麻素糖苷。
4.根据任一前述条目所述的基因修饰的宿主细胞,其中异戊二烯基供体选自香叶基二磷酸、橙花基二磷酸、法呢基二磷酸、二甲基烯丙基二磷酸和香叶基香叶基焦磷酸的组。
5.根据条目4所述的基因修饰的宿主细胞,其中异戊二烯基供体是香叶基二磷酸。
6.根据任一前述条目所述的基因修饰的宿主细胞,其中异戊二烯基受体是选自己酸、丁酸、戊酸、庚酸、辛酸、壬酸、癸酸、4-甲基己酸、5-己酸和6-庚酸的组的脂肪酸的衍生物。
7.根据条目6所述的基因修饰的宿主细胞,其中异戊二烯基受体选自橄榄酸、divarinolic acid、橄榄醇、呋罗苯异戊酮、白藜芦醇、柚皮素、间苯三酚和尿黑酸的组。
8.根据条目7所述的基因修饰的宿主细胞,其中异戊二烯基受体是橄榄酸和/或divarinolic acid。
9.根据任一前述条目的基因修饰的宿主细胞,其中大麻素受体和/或大麻素糖苷是人或动物大麻素受体的激动剂或拮抗剂。
10.根据条目9所述的基因修饰的宿主细胞,其中,大麻素受体和/或大麻素糖苷是非精神作用的或比THC低至少10%的精神作用。
11.根据前述任一条目所述的基因修饰的宿主细胞,其中大麻素受体是中性或酸性的。
12.根据前述任一条目所述的基因修饰的宿主细胞,其中大麻素受体选自以下的组:大麻色烯型(CBC)、大麻萜酚型(CBG)、大麻二酚型(CBD)、四氢大麻酚型(THC)、大麻环酚型(CBL)、大麻艾尔松型(CBE)、大麻酚型 (CBN)、脱氢大麻二酚型(CBND)和二羟基大麻酚型(CBT)。
13.根据条目12所述的基因修饰的宿主细胞,其中大麻素受体选自以下的组:大麻萜酚酸(CBGA)、大麻萜酚酸单甲基醚(CBGAM)、大麻萜酚单甲基醚(CBGM)、次大麻萜酚酸(CBGVA)、次大麻萜酚(CBGV)、大麻色烯酸 (CBCA)、次大麻色烯酸(CBCVA)、次大麻色烯(CBCV)、大麻二酚酸(CBDA)、大麻二酚单甲基醚(CBDM)、大麻二酚-C4(CBD-C4)、次大麻二酚酸 (CBDVA)、次大麻二酚(CBDV)、大麻二酚可尔(CBD-C1)、Δ9-反式四氢大麻酚(Δ9-THC)、Δ9-四氢大麻酚(Δ9-THC)、Δ9-顺式四氢大麻酚(Δ9-THC)、四氢大麻酚酸(THCA)、Δ9-四氢大麻酚酸A(THCA-A)、Δ9-四氢大麻酚酸 B(THCA-B)、Δ9-四氢大麻酚酸-C4(THCA-C4)、Δ9-四氢大麻酚 -C4(THC-C4)、Δ9-四氢次大麻酚酸(THCVA)、Δ9-四氢次大麻酚(THCV)、Δ9- 四氢大麻酚可尔酸(THCA-C1)、Δ9-四氢大麻酚可尔(THC-C1)、Δ7-顺式-异- 四氢次大麻酚、Δ8-四氢大麻酚酸(Δ8-THCA)、Δ8-反式-四氢大麻酚 (Δ8-THC)、Δ8-四氢大麻酚(Δ8-THC)、Δ8-顺式-四氢大麻酚(Δ8-THC)、大麻环酚酸(CBLA)、大麻环酚(CBL)、次大麻环酚(CBLV)、大麻艾尔松酸 A(CBEA-A)、大麻艾尔松酸B(CBEA-B)、大麻艾尔松(CBE)、cannabielsoinic acid、大麻二吡喃环烷、大麻二吡喃环烷酸、大麻酚酸(CBNA)、大麻酚甲基醚(CBNM)、大麻酚-C4(CBN-C4)、次大麻酚(CBV)、大麻酚-C2(CNB-C2)、大麻酚可尔(CBN-C1)、脱氢大麻二酚(CBND)、脱氢次大麻二酚(CBVD)、二羟基大麻酚(CBT)、10-乙氧基-9-羟基-δ-6a-四氢大麻酚、8,9-二羟基-δ-6a- 四氢大麻酚、二羟基次大麻酚(CBTVE)、脱氢大麻呋喃(DCBF)、大麻呋喃 (CBF)、大麻色酮(CBCN)、cannabiciuan(CBT)、10-氧亚基-δ-6a-四氢大麻酚 (OTHC)、δ-9-顺式-四氢大麻酚(顺式-THC)、3,4,5,6-四氢-7-羟基-α-α-2-三甲基-9-正丙基-2,6-桥亚甲基-2H-l-苯并氧杂环辛三烯-5-甲醇(OH-异-HHCV)、大麻利比索(CBR)、三羟基-δ-9-四氢大麻酚(triOH-THC)、perrottetinene、 perrottetinenic acid、11-Nor-9-羧基-THC、11-羟基-Δ9-THC、Nor-9-羧基-Δ9- 四氢大麻酚、tetrahydrocannabiphorol(THCP)、cannabidiphorol(CBDP)、 Cannabimovone(CBM)及其衍生物。
14.根据条目1至11所述的基因修饰的宿主细胞,其中大麻素受体是选自以下的组的内源性大麻素:花生四烯酰乙醇酰胺(anandamide,AEA)、2- 花生四烯酰乙醇酰胺(2-AG)、1-花生四烯酰乙醇酰胺(1-AG)和二十二碳六烯酰乙醇酰胺(DHEA,synaptamide)、油酰乙醇酰胺(OEA)、二十碳五烯酰乙醇酰胺、前列腺素乙醇酰胺、二十二碳六烯酰乙醇酰胺、亚麻酰乙醇酰胺、 5(Z),8(Z),11(Z)-二十碳三烯酸乙醇酰胺(米德酸乙醇酰胺)、十七烷酰乙醇酰胺、硬脂酰乙醇酰胺、二十二碳烯酰乙醇酰胺、神经酰基乙醇酰胺、二十三酰乙醇酰胺、木蜡酰乙醇酰胺、肉豆蔻酰乙醇酰胺、十五烷酰乙醇酰胺、棕榈油酰乙醇酰胺、二十二碳六烯酸(DHA)。
15.根据前述任一条目所述的基因修饰的宿主细胞,其中糖基供体选自 NTP-糖苷、NDP-糖苷和NMP-糖苷中的一种或多种。
16.根据条目15所述的基因修饰的宿主细胞,其中核苷酸糖苷的核苷选自尿苷、腺苷、鸟苷、胞苷和脱氧胸苷。
17.根据条目16所述的基因修饰的宿主细胞,其中糖基供体选自UDP- 糖苷、ADP-糖苷、CDP-糖苷、CMP-糖苷、dTDP-糖苷和GDP-糖苷。
18.根据条目17所述的基因修饰的宿主细胞,其中糖基供体选自 UDP-D-葡萄糖(UDP-Glc);UDP-半乳糖(UDP-Gal);UDP-D-木糖(UDP-Xyl);UDP-N-乙酰-D-葡糖胺(UDP-GlcNAc);UDP-N-乙酰-D-半乳糖胺 (UDP-GalNAc);UDP-D-葡糖醛酸(UDP-GlcA);UDP-D-呋喃半乳糖 (UDP-Galf);UDP-阿拉伯糖;UDP-鼠李糖;UDP-芹菜糖;UDP-2-乙酰胺基-2-脱氧-α-D-甘露糖醛酸酯;UDP-N-乙酰-D-半乳糖胺4-硫酸盐;UDP-N- 乙酰-D-甘露糖胺;UDP-2,3-双(3-羟基十四烷酰基)-葡糖胺;UDP-4-脱氧-4- 甲酰胺基-β-L-阿拉伯吡喃糖;UDP-2,4-双(乙酰胺基)-2,4,6-三脱氧-α-D-吡喃葡萄糖;UDP-半乳糖醛酸酯;UDP-3-氨基-3-脱氧-α-D-葡萄糖;鸟苷二磷酸-D-甘露糖(GDP-Man);鸟苷二磷酸-L-岩藻糖(GDP-Fuc);鸟苷二磷酸-L- 鼠李糖(GDP-Rha);胞苷单磷酸-N-乙酰神经氨酸(CMP-Neu5Ac);胞苷单磷酸-2-酮-3-脱氧-D-甘露辛酸(CMP-Kdo);和ADP-葡萄糖。
19.根据前述任一条目所述的基因修饰的宿主细胞,其中糖基转移酶源自植物或真菌。
20.根据条目19所述的基因修饰的宿主细胞,其中植物选自水稻、番红花、烟草、甜叶菊、本氏烟草(Nicotiana benthatamiana)和拟南芥。
21.根据条目1至20所述的基因修饰的宿主细胞,其中糖基转移酶能够使用选自NTP-糖苷、NDP-糖苷和/或NMP-糖苷的核苷酸糖苷作为糖基供体用于使大麻素糖基化。
22.根据条目21所述的基因修饰的宿主细胞,其中核苷酸糖苷的核苷选自尿苷、腺苷、鸟苷、胞苷和脱氧胸苷。
23.根据条目22所述的基因修饰的宿主细胞,其中糖基供体选自UDP- 糖苷、ADP-糖苷、CDP-糖苷、CMP-糖苷、dTDP-糖苷和GDP-糖苷。
24.根据前述任一条目所述的基因修饰的宿主细胞,其中糖基转移酶是 O-糖苷转移酶和/或C-糖苷转移酶。
25.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-糖基转移酶。
26.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素糖苷O-糖基转移酶。
27.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-葡糖基转移酶。
28.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-鼠李糖基转移酶。
29.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-木糖基转移酶。
30.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-阿拉伯糖基转移酶。
31.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-N-乙酰氨基半乳糖基转移酶。
32.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-N-乙酰氨基葡糖基转移酶。
33.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元/糖苷单-O-糖基转移酶。
34.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元/糖苷二-O-糖基转移酶。
35.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元/糖苷三-O-糖基转移酶。
36.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元/糖苷四-O-糖基转移酶。
37.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素 O-半乳糖基转移酶。
38.根据条目24所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素 O-葡糖醛酸基转移酶。
39.根据前述任一条目所述的基因修饰的宿主细胞,其中糖基转移酶选自EC2.4.1.-和EC2.4.2.-。
40.根据条目39所述的基因修饰的宿主细胞,其中糖基转移酶选自 EC2.4.1.17、EC2.4.1.35、EC2.4.1.159、EC2.4.1.203、EC2.4.1.234、EC2.4.1.236 和EC2.4.1.294。
41.根据条目39所述的基因修饰的宿主细胞,其中糖基转移酶选自 EC2.4.2.40。
42.根据前述任一条目所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-糖基转移酶和/或大麻素糖苷O-糖基转移酶,任选地与SEQ ID NO:1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、 33、35、37、39、101、103、105、107、109、111、113、115、117、119、 121、123、125、127、129、131、133、135、137、139、141、143、145、 147、149、151、153、155、157、159、161、163、165、167、169、171、 173、175、177、179、181、183、185、187、189、191、193、195、197、 199、201、203、205或207中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-糖基转移酶和/或大麻素糖苷O-糖基转移酶。
43.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:107、109、111、113、117、119、121、125、127、129、131、133、135、 137、139、141、143、147、149、151、153、155、157、159、161、163、 165、167、169、171、173、175、177、179、181、183、185、187、189、191、193、195、197、199、201、203、205、207中任一项包括的大麻素苷元O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少 90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
44.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素糖苷O-糖基转移酶,任选地与SEQ ID NO:115、123或145中任一项包括的大麻素糖O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素糖苷O-糖基转移酶。
45.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-葡糖基转移酶,任选地与SEQ ID NO:107、109、111、117、119、 121、125、127、129、131、133、135、137、139、141、143、147、149、 151、153、155、157、159、161、163、165、167、169、171、173、175、 177、179、181、183、185、187、189、191、193、195、197、199、201、 203、205或207中任一项包括的大麻素苷元O-葡糖基转移酶具有至少 70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-葡糖基转移酶。
46.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-鼠李糖基转移酶,任选地与SEQ ID NO:107、125、127、147、149、 151、157、159、161、177、183、191、197或207中任一项包括的大麻素苷元O-鼠李糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-鼠李糖基转移酶。
47.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-木糖基转移酶,任选地与SEQ ID NO:107、113、125、127、147、 149、151、157、159、161、177、183、191、197或207中任一项包括的大麻素苷元O-木糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-木糖基转移酶。
48.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-阿拉伯糖基转移酶,任选地与SEQ ID NO:107、125、127、147、 149、151、157、159、161、177、183、191、197或207中任一项包括的大麻素苷元O-阿拉伯糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-阿拉伯糖基转移酶。
49.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-N-乙酰氨基半乳糖基转移酶,任选地与SEQ ID NO:107、125、127、147、149、151、157、159、161、177、183、191、197或207中任一项包括的大麻素苷元O-N-乙酰氨基半乳糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元O-N-乙酰氨基半乳糖基转移酶。
50.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元O-N-乙酰氨基葡糖基转移酶,任选地与SEQ ID NO:107、125、127、 147、149、151、157、159、161、177、183、191、197或207中任一项包括的大麻素苷元O-N-乙酰氨基葡糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性的大麻素苷元O-N-乙酰氨基葡糖基转移酶。
51.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元/糖苷二-O-糖基转移酶,任选地与SEQ ID NO:107、115、123、125、 127、133、135、145、149、151、157、159、161、165、167、173、175、 177、185、191、195或207中任一项包括的大麻素苷元/糖苷二-O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元/糖苷二-O-糖基转移酶。
52.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是大麻素苷元/糖苷三-O-糖基转移酶,任选地与SEQ ID NO:107、115、123、145、 157、159、191或207中任一项包括的大麻素苷元/糖苷三-O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的大麻素苷元/糖苷三-O-糖基转移酶。
53.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶是四-O- 糖基转移酶,任选地与SEQ ID NO:207中任一项包括的大麻素苷元/糖苷四 -O-糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性的四-O-糖基转移酶。
54.根据条目42所述的基因修饰的宿主细胞,其中该糖基转移酶是糖基转移酶家族73。
55.根据条目54所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:107、157、159、191和/或207中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
56.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:135、143、147和/或171中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
57.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:107、109、111、113、117、125、127、129、135、137、139、141、147、 149、151、153、157、159、161、177、179、183、191、193、197、201、 205或207中任一项包括的糖基化CBD、CBDV和/或CBDA的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少 95%、诸如至少99%、诸如100%)同一性。
58.根据条目42所述的基因修饰的宿主细胞,,其中糖基转移酶与SEQ ID NO:107、109、119、125、127、135、137、147、149、151、157、159、 161、165、167、173、175、177、179、183、185、187、189、191、195、 201、205或207中任一项包括的糖基化CBG、CBGV和/或CBGA的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
59.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:107、111、117、121、125、127、131、143、149、155、157、159、163、 169、171、191、199、201、203或207中任一项包括的THC糖基化糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
60.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:125、127、133、135、149、151、157、159、175、177、181、191、 195或207中任一项包括的CBN糖基化糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
61.根据条目42所述的基因修饰的宿主细胞,其中该糖基转移酶与SEQ ID NO:107、125、127、135、149、151、157、159、175、177、191、201 或207中任一项包括的CBC糖基化糖基转移酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
62.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:SEQID NO:147、157、107、159、191、171、135、143中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
63.根据条目42至62所述的基因修饰的宿主细胞,其中序列同一性为至少90%,诸如至少95%、诸如至少99%、诸如100%。
64.根据条目63所述的基因修饰的宿主细胞,其中序列同一性为至少 99%,诸如100%。
65.根据条目42所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:25、27、29、31、33、35、37、39、101或103中包括的糖基转移酶具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性。
66.根据条目65所述的基因修饰的宿主细胞,其中糖基转移酶与SEQ ID NO:25、27、29、31、33、35、37、39、101或103中任一项包括的糖基转移酶具有至少95%(诸如至少99%、诸如100%)同一性。
67.根据条目66所述的基因修饰的宿主细胞,其中糖基转移酶是SEQ ID NO:25、27、29、31、33、35、37、39、101或103中任一项包括的糖基转移酶。
68.根据前述任一条目所述的基因修饰的宿主细胞,其中表达的糖基转移酶不存在用于分泌的靶向糖基转移酶的信号肽。
69.根据前述任一条目所述的基因修饰的宿主细胞,其中糖基转移酶催化糖基基团与大麻素苷元或大麻素糖苷之间的1,2-;1,3-;1,4-和/或1,6-糖苷键的形成。
70.根据条目69所述的基因修饰的宿主细胞,其中糖基转移酶催化糖基基团与大麻素苷元或大麻素糖苷之间1,4-和/或1,6-糖苷键的形成。
71.根据条目70所述的基因修饰的宿主细胞,其中糖基转移酶是SEQ ID NO:115中包括的糖基转移酶,并且催化糖基基团与大麻素苷元或大麻素糖苷之间1,4-糖苷键的形成。
72.根据条目70所述的基因修饰的宿主细胞,其中糖基转移酶是SEQ ID NO:145中包括的糖基转移酶,并且催化糖基基团与大麻素苷元或大麻素糖苷之间1,6-糖苷键的形成。
73.根据前述任一条目所述的基因修饰的宿主细胞,其中编码糖基转移酶的异源基因与SEQ ID NO:2、4、6、8、10、12、14、16、18、20、22、 24、26、28、30、32、34、36、38、40、102、104、106、108、110、112、 114、116、118、120、122、124、126、128、130、132、134、136、138、140、142、144、146、148、150、152、154、156、158、160、162、164、 166、168、170、172、174、176、178、180、182、184、186、188、190、 192、194、196、198、200、202、204、206或208中任一项包括的编码糖基转移酶的基因具有至少70%(诸如至少75%、诸如至少80%、诸如至少 90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
74.根据条目73所述的基因修饰的宿主细胞,其中编码糖基转移酶的异源基因与SEQ ID NO:148、158、108、160、192、172、137、144中任一项包括的糖基转移酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
75.根据条目73至74所述的基因修饰的宿主细胞,其中序列同一性为至少90%,诸如至少95%、诸如至少99%、诸如100%。
76.根据条目75所述的基因修饰的宿主细胞,其中序列同一性为至少 99%,诸如100%。
77.根据条目73项所述的基因修饰的宿主细胞,其中编码糖基转移酶的异源基因与SEQ ID NO:26、28、30、32、34、36、38、40、102或104中任一项包括的编码糖基转移酶的基因具有至少90%(诸如至少95%、诸如至少99%、诸如100%)同一性。
78.根据条目77所述的基因修饰的宿主细胞,其中编码糖基转移酶的异源基因与SEQ ID NO:26、28、30、32、34、36、38、40、102或104中任一项包括的编码糖基转移酶的基因具有至少95%(诸如至少99%、诸如100%) 同一性。
79.根据条目78所述的基因修饰的宿主细胞,其中编码糖基转移酶的异源基因是SEQ ID NO:26、28、30、32、34、36、38、40、102或104中任一项包括的编码糖基转移酶的基因。
80.根据前述任一条目所述的基因修饰的宿主细胞,其中大麻素糖苷的水溶解度比相应的未糖基化的大麻素高至少10%。
81.根据前述任一条目所述的基因修饰的宿主细胞,其中大麻素糖苷对 UV或热降解的抗性比相应的未糖基化的大麻素高至少10%。
82.根据前述任一条目所述的基因修饰的宿主细胞,其中当同等地给药于哺乳动物时,该大麻素糖苷的口服摄取量比相应的未糖基化的大麻素高至少10%。
83.根据前述任一条目所述的基因修饰的宿主细胞,其中当同等地给药于哺乳动物时,该大麻素糖苷的生物半衰期比相应的未糖基化的大麻素高至少10%。
84.根据前述任一条目所述的基因修饰的宿主细胞,其中当同等地给药于哺乳动物时,该大麻素糖苷在峰值浓度下的CNS浓度比相应的未糖基化的大麻素高至少10%。
85.根据前述任一条目所述的基因修饰的宿主细胞,其中与相应的未糖基化的大麻素相比,该大麻素糖苷的药代动力学提高至少10%,如通过溶解度测定、化学稳定性测定、Caco-2双向渗透性测定、肝微粒体清除测定和/或血浆稳定性测定所测量的。
86.根据前述任一条目所述的基因修饰的宿主细胞,其中与相应的未糖基化的大麻素相比,该大麻素糖苷在酸性水溶液中,任选地在pH值为0至 7(诸如pH值为0.5至4、诸如pH值为0.5至2、诸如pH值为约1)的溶液中的稳定性提高至少10%。
87.根据前述任一条目所述的基因修饰的宿主细胞,其中与相应的未糖基化的大麻素相比,该大麻素糖苷在碱性水溶液中,任选地在pH值为7至 14(诸如pH值为9至14、诸如pH值为10至13、诸如pH值为约12.5)的溶液中的稳定性提高至少10%。
88.根据前述任一条目所述的基因修饰的宿主细胞,其中与相应的未糖基化的大麻素相比,该大麻素糖苷在水溶液中(任选地在具有至少8mg/L O2(诸如至少20mg/L O2、诸如至少40mg/L O2、诸如至少80mg/L O2的溶液,诸如用O2饱和的溶液中)的抗氧化性提高至少10%。
89.根据前述任一条目所述的基因修饰的宿主细胞,其中与相应的未糖基化的大麻素相比,该大麻素糖苷对基因修饰的宿主细胞的毒性降低至少 10%,任选地具有比相应的未糖基化的大麻素低至少10%(诸如低至少25%、诸如低至少75%、诸如低至少100%)的LC50。
90.根据前述任一条目所述的基因修饰的宿主细胞,其中该大麻素糖苷是C-糖苷或O-糖苷或其衍生物或组合。
91.根据前述任一条目所述的基因修饰的宿主细胞,其中该大麻素糖苷是选自以下的糖苷:大麻色烯型(CBC)、大麻萜酚型(CBG)、大麻二酚型 (CBD)、四氢大麻酚型(THC)、大麻环酚型(CBL)、大麻艾尔松型(CBE)、大麻酚型(CBN)、脱氢大麻二酚型(CBND)和二羟基大麻酚型。
92.根据条目91项所述的基因修饰的宿主细胞,其中该大麻素糖苷选自以下的糖苷:大麻二酚(CBD)、大麻二酚酸(CBDA)、次大麻二酚(CBDV)、四氢大麻酚(THC)、四氢大麻酚酸(THCA)、四氢次大麻酚(THCV)、次大麻色烯(CBCV)、大麻萜酚(CBG)、大麻酚(CBN)、11-nor-9-羧基-THC和Δ8- 四氢大麻酚。
93.根据前述任一条目所述的基因修饰的宿主细胞,其中该大麻素糖苷包括与选自以下的糖共价连接的大麻素苷元或大麻素糖苷:木糖;鼠李糖;半乳糖;N-乙酰葡糖胺;N-乙酰半乳糖胺;和阿拉伯糖。
94.根据前述任一条目所述的基因修饰的宿主细胞,其中该大麻素糖苷选自大麻素-1'-O-β-D-糖苷、大麻素-1'-O-β-糖基-3'-O-β-糖苷和大麻素 -3'-O-β-D-糖苷。
95.根据条目93所述的基因修饰的宿主细胞,其中,该大麻素糖苷选自 CBD-1'-O-β-D-糖苷、CBD-1’-O-β-糖基-3’-O-β-糖苷、CBDV-1’-O-β-D-糖苷、 CBDV-1’-O-β-糖基-3’-O-β-糖苷、CBG-1'-O-β-D-糖苷、CBG-1’-O-β-糖基 -3’-O-β-糖苷、THC-1'-O-β-D-糖苷、CBN-1'-O-β-D-糖苷、11-nor-9-羧基 -THC-1’-O-β-D-糖苷、CBDA-3’-O-β-D-糖苷和CBC-3’-O-β-D-糖苷。
96.根据前述任一条目所述的基因修饰的宿主细胞,其中该大麻素糖苷选自大麻素葡糖苷;大麻素葡糖醛酸苷;大麻素木糖苷;大麻素鼠李糖苷;大麻素半乳糖苷;大麻素N-乙酰氨基葡糖苷;大麻素N-乙酰氨基半乳糖苷和大麻素阿拉伯糖苷。
97.根据条目96所述的基因修饰的宿主细胞,其中,大麻素糖苷选自大麻素-1'-O-β-D-葡糖苷;大麻素-1'-O-β-D-葡糖醛酸苷;大麻素-1'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖苷;大麻素-1'-O-β-D-半乳糖苷;大麻素 -1'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖苷;大麻素 -1'-O-β-D-N-乙酰半乳糖胺;大麻素-1'-O-β-D-纤维二糖苷;大麻素-1'-O-β-D- 龙胆二糖苷;大麻素-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷;大麻素-1'-O-β-D- 葡糖醛酸基-3'-O-β-D-葡糖醛酸苷;大麻素-1'-O-β-D-木糖基-3'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖基-3'-O-β-D-鼠李糖苷;大麻素-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;和大麻素 -1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺。
98.根据条目97所述的基因修饰的宿主细胞,其中,该大麻素糖苷选自 CBD-1'-O-β-D-纤维二糖苷;CBD-1'-O-β-D-龙胆二糖苷;CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷;CBD-1'-O-β-D-葡萄糖醛糖基-3'-O-β-D-葡萄糖醛糖苷;CBD-1'-O-β-D-木糖基-3'-O-β-D-木糖苷CBD-1'-O-α-L-鼠李糖基 -3'-O-α-L-鼠李糖苷;CBD-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷; CBD-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;CBD-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;CBD-1'-O-β-D-N-乙酰半乳糖胺 -3'-O-β-D-N-乙酰半乳糖胺;CBDV-1'-O-β-D-纤维二糖苷;CBDV-1'-O-β-D- 龙胆二糖苷;CBDV-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷;CBDV-1'-O-β-D-葡萄糖醛糖基-3'-O-β-D-葡萄糖醛糖苷;CBDV-1'-O-β-D-木糖基-3'-O-β-D-木糖苷;CBDV-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷;CBDV-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBDV-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;CBDV-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷; CBDV-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺;CBG-1'-O-β-D- 纤维二糖苷;CBG-1'-O-β-D-龙胆二糖苷;CBG-1'-O-β-D-葡糖基-3'-O-β-D- 葡糖苷;CBG-1'-O-β-D-葡萄糖醛糖基-3'-O-β-D-葡萄糖醛糖苷; CBG-1'-O-β-D-木糖基-3'-O-β-D-木糖苷CBG-1'-O-α-L-鼠李糖基-3'-O-α-L- 鼠李糖苷;CBG-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBG-1'-O-β-D-N- 乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;CBG-1'-O-β-D-阿拉伯糖基 -3'-O-β-D-阿拉伯糖苷;CBG-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺;THC-1'-O-β-D-葡糖苷;THC-1'-O-β-D-纤维二糖苷;THC-1'-O-β-D- 龙胆二糖苷;THC-1'-O-β-D-葡萄糖醛糖苷;THC-1'-O-β-D-木糖苷; THC-1'-O-α-L-鼠李糖苷;THC-1'-O-β-D-半乳糖苷;THC-1'-O-β-D-N-乙酰氨基葡糖苷;THC-1'-O-β-D-阿拉伯糖苷;THC-1'-O-β-D-N-乙酰氨基半乳糖苷;CBN-1'-O-β-D-葡糖苷;CBN-1'-O-β-D-纤维二糖苷;CBN-1'-O-β-D-龙胆二糖苷;CBN-1'-O-β-D-葡萄糖醛糖苷;CBN-1'-O-β-D-木糖苷;CBN-1'-O-α-L- 鼠李糖苷;CBN-1'-O-β-D-半乳糖苷;CBN-1'-O-β-D-N-乙酰氨基葡糖苷; CBN-1'-O-β-D-阿拉伯糖苷;CBN-1'-O-β-D-N-乙酰氨基半乳糖苷; CBDA-1'-O-β-D-葡糖苷;CBDA-1'-O-β-D-纤维二糖苷;CBDA-1'-O-β-D-龙胆二糖苷;CBDA-1'-O-β-D-葡萄糖醛糖苷;CBDA-1'-O-β-D-木糖苷; CBDA-1'-O-α-L-鼠李糖苷;CBDA-1'-O-β-D-半乳糖苷;CBDA-1'-O-β-D-N- 乙酰氨基葡糖苷;CBDA-1'-O-β-D-阿拉伯糖苷;CBDA-1'-O-β-D-N-乙酰氨基半乳糖苷;CBC-1'-O-β-D-葡糖苷;CBC-1'-O-β-D-纤维二糖苷; CBC-1'-O-β-D-龙胆二糖苷;CBC-1'-O-β-D-葡萄糖醛糖苷;CBC-1'-O-β-D-木糖苷;CBC-1'-O-α-L-鼠李糖苷;CBC-1'-O-β-D-半乳糖苷;CBC-1'-O-β-D-N- 乙酰氨基葡糖苷;CBC-1'-O-β-D-阿拉伯糖苷;and CBC-1'-O-β-D-N-乙酰氨基半乳糖苷。
99.根据前述任一条目所述的基因修饰的宿主细胞,进一步包括能够产生该大麻素受体的起作用的生物合成代谢途径,其中该途径包括选自以下的一种或多种多肽:
a)乙酰乙酰-CoA硫解酶(ACT),所述ACT将乙酰-CoA前体转化为乙酰乙酰-CoA;
b)HMG-CoA合酶(HCS),所述HCS将乙酰乙酰-CoA前体转化为 HMG-CoA;
c)HMG-CoA还原酶(HCR),所述HCR将HMG-CoA前体转化为甲羟戊酸;
d)甲羟戊酸激酶(MVK),所述NVK将甲羟戊酸前体转化为甲羟戊酸-5- 磷酸;
e)磷酸甲羟戊酸激酶(PMK),所述PMK将甲羟戊酸-5-磷酸前体转化为甲羟戊酸二磷酸;
f)甲羟戊酸焦磷酸脱羧酶(MPC),所述MPC将甲羟戊酸二磷酸前体转化为异戊烯基二磷酸(IPP);
g)异戊烯基二磷酸/二甲基烯丙基二磷酸异构酶(IPI),所述IPI将IPP前体转化为二甲基烯丙基二磷酸(DMAPP);
h)香叶基二磷酸合酶(GPPS),所述GPPS将IPP和DMAPP缩合成香叶基二磷酸(GPP);
i)酰基活化酶(AAE),所述AAE将脂肪酸前体转化为脂肪酰基-COA;
j)3,5,7-三氧亚基十二烷酰基-CoA合酶(TKS),所述TKS将脂肪酸-CoA 前体转化为3,5,7-三氧亚基十一烷酰基-CoA;
k)橄榄酸环化酶(OAC),所述OAC将3,5,7-三氧亚基十一烷酰基-CoA 前体转化为divarinolic acid;
l)橄榄酸环化酶(OAC),所述OAC将3,5,7-三氧亚基十二烷酰基-CoA 前体转化为橄榄酸;
m)TKS-OAC融合酶,所述TKS-OAC融合酶将脂肪酸-CoA前体转化为3,5,7-三氧亚基十一烷酰基-CoA、将3,5,7-三氧亚基十一烷酰基-CoA前体转化为divarinolic acid和将3,5,7-三氧亚基十二烷酰基-CoA前体转化为橄榄酸;
n)大麻萜酚酸合酶(CBGAS),所述CBGAS将GPP和橄榄酸缩合为大麻萜酚酸(CBGA);
o)大麻萜酚酸合酶(CBGAS),所述CBGAS将GPP和divarinolic acid缩合为次大麻萜酚酸(CBGVA);
p)大麻二酚酸合酶(CBDAS),所述CBDAS分别将CBGA酸和/或 CBGVA转化为大麻二酚酸(CBDA)和/或次大麻二酚酸(CBDVA);
q)四氢大麻酚酸合酶(THCAS),所述THCAS分别将CBGA和/或 CBGVA转化为四氢大麻酚酸(THCA)和/或四氢次大麻酚酸(THCVA);
r)大麻色烯酸合酶(CBCAS),所述CBCAS分别将CBGA和/或CBGVA 转化为大麻色烯酸(CBCA)和/或次大麻色烯酸(CBCVA);
s)核苷酸-葡萄糖合酶,所述核苷酸-葡萄糖合酶将蔗糖和核苷酸转化为果糖和核苷酸-葡萄糖;
t)核苷酸-半乳糖4-差向异构酶,所述核苷酸-半乳糖4-差向异构酶将核苷酸-葡萄糖转化为核苷酸-半乳糖;
u)核苷酸-(葡糖醛酸)-脱羧酶,该核苷酸-(葡萄醛酸)-脱羧酶将核苷酸- 葡糖醛酸转化为核苷酸-木糖;
v)核苷酸-4-酮-6-脱氧-葡萄糖3,5-差向异构酶和核苷酸-4-酮-鼠李糖4- 酮-还原酶,它们一起将核苷酸-4-酮-6-脱氧-葡萄糖和NADPH转化为核苷酸-鼠李糖和NADP+;
w)核苷酸-葡萄糖4,6-脱水酶,该核苷酸-葡萄糖4,6-脱水酶将核苷酸- 葡萄糖和NAD化为核苷酸-4-酮-6-脱氧-葡萄糖和NADH;
x)核苷酸-葡萄糖4,6-脱水酶和核苷酸-4-酮-6-脱氧-葡萄糖3,5差向异构酶以及核苷酸-4-酮-鼠李糖-4-酮-还原酶,它们一起将核苷酸-葡萄糖和 NAD+以及NADPH转化为核苷酸-鼠李糖+NADH+NADP+;
y)核苷酸-葡萄糖6脱氢酶,该核苷酸-葡萄糖6脱氢酶将核苷酸-葡萄糖和2NAD+转化为核苷酸-葡糖醛酸和2NADH;
z)核苷酸-阿拉伯糖4-差向异构酶,该核苷酸-阿拉伯糖4-差向异构酶将核苷酸-木糖转化为核苷酸-阿拉伯糖;以及
aa)核苷酸-N-乙酰葡糖胺4-差向异构酶,该核苷酸-N-乙酰葡糖胺4- 差向异构酶将核苷酸-N-乙酰葡糖胺转化为核苷酸-N-乙酰半乳糖胺。
100.根据条目99所述的基因修饰的宿主细胞,其中:
a)ACT与酿酒酵母中的天然Erg10具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
b)HCS与酿酒酵母中的天然Erg13具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
c)HCS与酿酒酵母中的天然HMG1或HMG2具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
d)MVK与酿酒酵母中的天然Erg12具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
e)PMK与酿酒酵母中的天然Erg8具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
f)MPC与酿酒酵母中的天然MVD1具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
g)IPI与酿酒酵母中的天然IDI1具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
h)GPPS与SEQ ID NO:45或229中包括的GPPS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
i)AAE与SEQ ID NO:47或239中包括的AAE具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
j)TKS与SEQ ID NO:49中包括的TKS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
k)OAC与SEQ ID NO:51中包括的OAC具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%) 同一性;
l)TKS-OAC融合酶与SEQ ID NO:227中包括的TKS-OAC融合酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
m)CBGAS与SEQ ID NO:53、235、237中包括的CBGAS具有至少 70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
n)CBDAS与SEQ ID NO:57或233中包括的CBDAS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
o)THCAS与SEQ ID NO:55或231中包括的THCAS具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
p)CBCAS与SEQ ID NO:59中包括的CBCAS具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
q)核苷酸-葡萄糖合酶是UDP-葡萄糖合酶并且与SEQ ID NO:209中包括的UDP-葡萄糖合酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
r)核苷酸-半乳糖4-差向异构酶是UDP-半乳糖4-差向异构酶并且与 SEQ ID NO:211中包括的UDP-半乳糖4-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
s)核苷酸-(葡糖醛酸)-脱羧酶是UDP-葡糖醛酸脱羧酶并且与SEQ ID NO:213中包括的UDP-葡糖醛酸脱羧酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
t)核苷酸-4-酮-6-脱氧-葡萄糖3,5-差向异构酶是UDP-4-酮-6-脱氧-葡萄糖3,5-差向异构酶并且与SEQ ID NO:215或219中包括的UDP-4-酮-6-脱氧 -葡萄糖3,5-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
u)核苷酸-4-酮-鼠李糖-4-酮还原酶是UDP-4-酮-鼠李糖-4-酮还原酶并且与SEQID NO:215或219中包括的UDP-4-酮-鼠李糖-4-酮还原酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
v)核苷酸-葡萄糖4,6脱水酶是UDP-葡萄糖4,6-脱水酶并且与SEQ ID NO:217或219中包括的UDP-葡萄糖4,6-脱水酶具有至少70%(诸如至少 75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
w)核苷酸-葡萄糖6脱氢酶是UDP-葡萄糖6-脱氢酶并且与SEQ ID NO: 221中包括的UDP-葡萄糖6-脱氢酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;
x)核苷酸-阿拉伯糖4-差向异构酶是UDP-阿拉伯糖4-差向异构酶并且与SEQ IDNO:223中包括的UDP-阿拉伯糖4-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性;以及
y)核苷酸-N-乙酰葡糖胺4-差向异构酶是UDP-N-乙酰葡糖胺4-差向异构酶并且与SEQ ID NO:225中包括的UDP-N-乙酰葡糖胺4-差向异构酶具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
101.根据条目100所述的基因修饰的宿主细胞,其中:
a)ACT是酿酒酵母中的天然Erg10;
b)HCS是酿酒酵母中的天然Erg13;
c)HCR是酿酒酵母中的天然HMG1;
d)HCR是酿酒酵母中的天然HMG2;
e)MVK是酿酒酵母中的天然Erg12;
f)PMK是酿酒酵母中的天然Erg8;
g)MPC是酿酒酵母中的天然MVD1;
h)IPI是酿酒酵母中的天然IDI1;
i)GPPS是SEQ ID NO:45或229的GPPS;
j)AAE是SEQ ID NO:47或238的AAE;
k)TKS是SEQ ID NO:49的TKS;
l)OAC是SEQ ID NO:51的OAC;
m)TKS-OAC融合酶是SEQ ID NO 227中包括的TKS-OAC融合酶
n)CBGAS是SEQ ID NO:53、235或237的CBGAS;
o)CBDAS是SEQ ID NO:57或233的CBDAS;
p)THCAS是SEQ ID NO:55或231的THCAS;
q)CBCAS是SEQ ID NO:59的CBCAS;
r)UDP-葡萄糖合酶是SEQ ID NO:209中包括的UDP-葡萄糖合酶;
s)UDP-半乳糖4-差向异构酶是SEQ ID NO:211中包括的UDP-半乳糖 4-差向异构酶;
t)UDP-葡糖醛酸脱羧酶是SEQ ID NO:213中包括的UDP-葡糖醛酸脱羧酶;
u)UDP-4-酮-6-脱氧-葡萄糖3,5-差向异构酶是SEQ ID NO:215或219中包括的UDP-4-酮-6-脱氧-葡萄糖3,5-差向异构酶;
v)UDP-4-酮-鼠李糖-4-酮还原酶是SEQ ID NO:215或219中包括的UDP-4-酮-鼠李糖-4-酮还原酶;
w)UDP-葡萄糖4,6-脱水酶是SEQ ID NO:217或219中包括的UDP-葡萄糖4,6-脱水酶;
x)UDP-葡萄糖6-脱氢酶是SEQ ID NO:221中包括的UDP-葡萄糖6-脱氢酶;
y)UDP-阿拉伯糖4-差向异构酶是SEQ ID NO:223中包括的UDP-阿拉伯糖4-差向异构酶;以及
z)UDP-N-乙酰葡糖胺4-差向异构酶是SEQ ID NO:225中包括的UDP-N-乙酰葡糖胺4-差向异构酶。
102.根据前述任一条目所述的基因修饰的宿主细胞,其中起作用的生物合成代谢途径中包括的多个多肽对于基因修饰的宿主细胞是异源的。
103.根据前述任一条目所述的基因修饰的宿主细胞,其中基因修饰的宿主细胞进一步被基因修饰以提供增加量的用于起作用的生物合成代谢途径的至少一种多肽的底物。
104.根据前述任一条目所述的基因修饰的宿主细胞,其中经基因修饰的宿主细胞进一步被基因修饰以对来自起作用的生物合成代谢途径的一种或多种底物、中间体或产物分子表现出增加的耐受性。
105.根据前述任一条目所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞进一步被基因修饰以包括促进细胞内形成的大麻素糖苷的分泌的转运蛋白多肽。
106.根据前述任一条目所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是真核细胞、原核细胞或古生物细胞。
107.根据条目106所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是选自哺乳动物、昆虫、植物或真菌细胞的组的真核细胞。
108.根据条目107所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是大麻属、葎草属或甜菊属的植物细胞。
109.根据条目107所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是真菌宿主细胞,其选自由以下组成的门:子囊菌门、担子菌门、新丽鞭毛菌门、球囊菌门、芽枝霉门、壶菌门、接合菌门、卵菌门和微孢子菌门。
110.根据条目109所述的基因修饰的宿主细胞,其中基因修饰的真菌宿主细胞是选自由以下项组成的组的酵母:产子囊孢子酵母(内孢霉目)、产担孢子酵母和半知菌酵母(芽孢纲)。
111.根据条目110所述的基因修饰的宿主细胞,其中基因修饰的酵母宿主细胞选自由以下项组成的属:酵母属、克鲁维酵母菌属、念珠菌属、毕赤酵母属、德巴利氏酵母属、汉逊酵母属、耶氏酵母属、接合酵母属和裂殖酵母属。
112.根据条目111所述的基因修饰的宿主细胞,其中基因修饰的宿主细胞选自由以下组成的种:乳酸克鲁维酵母、卡尔酵母、酿酒酵母、糖化酵母、道格拉斯酵母、克鲁维酵母、诺地酵母、卵形酵母、布拉酵母和解脂耶氏酵母。
113.根据条目109所述的基因修饰的宿主细胞,其中基因修饰的真菌宿主细胞是丝状真菌。
114.根据条目113所述的基因修饰的宿主细胞,其中丝状真菌基因修饰的宿主细胞选自子囊菌门、真菌门和卵菌门。
115.根据条目114所述的基因修饰的宿主细胞,其中丝状真菌宿主细胞选自由以下组成的组的属:支顶孢属、曲霉属、短柄霉属、烟管菌属、拟蜡菌属、金孢子菌属、鬼伞属、Corio/us、隐球菌属、Filibasidium、镰刀菌属、腐质霉属、稻瘟菌属、毛霉属、毁丝霉属、新美鞭菌属、链孢霉、拟青霉属、青霉菌属、毛平革菌、白腐菌属、瘤胃壶菌属、侧耳属、裂褶菌属、篮状菌属、热子囊菌属、梭孢壳属、弯颈霉属、栓菌属和木霉属。
116.根据条目115所述的基因修饰的宿主细胞,其中丝状真菌宿主细胞选自由以下组成的的种:泡盛曲霉(Aspergillus awamori)、臭曲霉(Aspergillus foetidus)、烟曲霉(Aspergillus fumigatus)、日本曲霉(Aspergillus japonicus)、构巢曲霉(Aspergillus nidulans)、黑曲霉(Aspergillus niger)、米曲霉(Aspergillus oryzae)、烟管菌(Bjerkandera adusta)、干拟蜡菌(Ceriporiopsis aneirina)、 Ceriporiopsiscaregiea、Ceriporiopsis gilvescens、潘诺希塔拟蜡菌 (Ceriporiopsis pannocinta)、环带拟蜡菌(Ceriporiopsis rivulosa)、微红拟蜡菌 (Ceriporiopsis subrufa)、虫拟蜡菌(Ceriporiopsis subvermispora)、狭边金孢子菌(Chrysosporiuminops)、嗜角质金孢子菌(Chrysosporiumkeratinophilum)、卢克诺文思金孢子菌(Chrysosporiumlucknowense)、类状金孢子菌 (Chrysosporium merdarium)、毡金孢子菌(Chrysosporiumpannicola)、昆士兰金孢子菌(Chrysosporium queenslandicum)、热带金孢子菌(Chrysosporium tropicum)、褐薄金孢子菌(Chrysosporium zonatum)、灰盖鬼伞菌(Coprinus cinereus)、毛云芝菌(Coriolus hirsutus)、Fusarium bactridioides、禾谷镰孢菌 (Fusarium cerealis)、克鲁克威尔镰孢菌(Fusarium crookwellense)、黄色镰孢菌(Fusarium culmorum)、禾谷镰刀菌(Fusarium graminearum)、禾赤镰孢菌 (Fusariumgraminum)、异孢镰刀菌(Fusarium heterosporum)、合欢木镰孢菌 (Fusarium negundi)、尖孢镰刀菌(Fusarium oxysporum)、多枝镰孢菌(Fusarium reticulatum)、粉红镰孢菌(Fusarium roseum)、接骨木镰孢菌(Fusarium sambucinum)、肤色镰孢菌(Fusariumsarcochroum)、拟分枝镰孢菌(Fusarium sporotrichioides)、硫色镰孢菌(Fusariumsulphureum)、Fusarium torulosum、拟丝孢镰孢菌(Fusarium trichothecioides)、Fusarium venenatum、特异腐质霉 (Humicola insolens)、柔毛腐质霉(Humicolalanuginosa)、米黑毛霉(Mucor miehei)、嗜热毁丝霉(Myceliophthora thermophila)、粉色面包霉菌(Neurospora crassa)、产紫青霉(Penicillium purpurogenum)、黄孢原毛平革菌(Phanerochaete chrysosporium)、射脉齿菌(Phlebia radiata)、刺芹侧耳(Pleurotuseryngii)、土生梭孢壳霉(Thielavia terrestris)、长绒毛栓菌(Trametes villosa)、变色栓菌 (Trametes versicolor)、哈茨木霉(Trichoderma harzianum)、康宁木霉(Trichoderma koningii)、长枝木霉(Trichoderma longibrachiatum)、里氏木霉(Trichoderma reesei)和绿色木霉(Trichoderma viride)。
117.根据条目106所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是原核细胞。
118.根据条目117所述的基因修饰的宿主细胞,其中该原核细胞是大肠杆菌。
119.根据条目106所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是古生物细胞。
120.根据条目119所述的基因修饰的宿主细胞,其中该古生物细胞是藻类。
121.一种包括编码任一前述项的糖基转移酶的多核苷酸序列的多核苷酸构建体,其可操作地连接至与糖基编码多核苷酸异源的一个或多个控制序列。
122.根据条目121所述的多核苷酸构建体,其中编码糖基转移酶的多核苷酸与SEQID NO:2、4、6、8、10、12、14、16、18、20、22、24、26、 28、30、32、34、36、38、40、102、104、106、108、110、112、114、116、 118、120、122、124、126、128、130、132、134、136、138、140、142、144、146、148、150、152、154、156、158、160、162、164、166、168、 170、172、174、176、178、180、182、184、186、188、190、192、194、 196、198、200、202、204、206或208中任一项包括的编码糖基转移酶的基因具有至少70%(诸如至少75%、诸如至少80%、诸如至少90%、诸如至少95%、诸如至少99%、诸如100%)同一性。
123.一种表达载体,包括条目121或122所述的多核苷酸构建体。
124.一种基因修饰的宿主细胞,包括条目123所述的多核苷酸构建体或载体。
125.根据前述任一条目所述的基因修饰的宿主细胞,包括编码糖基转移酶和/或任何途径酶的基因的至少两个拷贝。
126.根据前述任一条目所述的基因修饰的宿主细胞,其中一个或多个天然基因被减毒、破坏和/或缺失。
127.根据前述任一条目所述的基因修饰的宿主细胞,其中该基因修饰的宿主细胞是通过减毒、破坏和/或缺失SGD ID SGD:S000005979的PDR12 来修饰的酿酒酵母菌株。
128.一种细胞培养物,包括根据前述任一条目所述的基因修饰的宿主细胞和生长培养基。
129.一种产生大麻素糖苷的方法,包括:
a)在允许基因修饰的宿主细胞产生大麻素糖苷的条件下培养根据条目 128所述的细胞培养物;和
b)任选地回收和/或分离所述大麻素糖苷。
130.根据条目129所述的方法,进一步包括选自以下的一个或多个要素:
a)在营养生长培养基中培养细胞培养物;
b)在需氧或厌氧条件下培养细胞培养物
c)在搅拌下培养细胞培养物;
d)在25至50℃的温度下培养细胞培养物;
e)在3-9之间的pH下培养细胞培养物;
f)将细胞培养物培养10小时至30天;以及
g)在分批补料、重复分批补料或半连续条件下培养细胞
h)在有机溶剂存在下培养细胞培养物以提高大麻素苷元的溶解度。
131.根据条目129至130所述的方法,进一步包括大麻素受体和/或大麻素糖苷的非酶促脱羧步骤。
132.根据条目131所述的方法,其中脱羧通过热处理、UV处理或碱度处理或其组合实现。
133.根据条目129至132所述的方法,进一步包括将一种或多种外源大麻素受体和/或核苷酸-糖苷补料至细胞培养物。
134.根据条目129至133所述的方法,其中回收和/或分离步骤包括将基因修饰的宿主细胞或细胞培养物的液相与基因修饰的宿主细胞或细胞培养物的固相分离以通过选自以下的一个或多个步骤获得包括大麻素糖苷的上清液:
a)分解基因修饰的宿主细胞以将细胞内的大麻素糖苷释放到上清液中;
b)使上清液与一种或多种吸附树脂接触以获得产生的大麻素糖苷的至少一部分;
c)使上清液与一个或多个离子交换或反相色谱柱接触以获得大麻素糖苷的至少一部分;和
d)结晶或提取大麻素糖苷;以及
e)蒸发液相溶剂以浓缩或沉淀大麻素糖苷;
从而回收和/或分离大麻素糖苷。
135.根据条目129至134所述的方法,其中大麻素糖苷的产率比通过来自甜叶菊的UGT76G1生产的产率高至少10%,诸如至少50%、诸如至少 100%、诸如至少150%、诸如至少200%。
136.根据138所述的方法,其中糖基化在体外进行。
137.根据条目129至136所述的方法,包括以下步骤:将大麻素糖苷加工成药用大麻素制剂,其包括向包括非植物细胞的条目128的细胞培养物补料在生长培养基中的起始材料;从细胞培养物中产生药用大麻素化合物以产生包含细胞培养物、生长培养基和药用大麻素化合物的混合物;对药物大麻素化合物进行处理,其中处理包括:使用选自由沉降、过滤和离心组成的组中的至少一种方法分离基因修饰的细胞;以及产生包括药用大麻素的药用大麻素制剂,其中混合物不含可检测量的植物杂质,该植物杂质选自由以下组成的组:多糖、木质素、色素、类黄酮、菲类、胶乳、树胶、树脂、蜡、杀虫剂、杀真菌剂、除草剂和花粉。
138.一种用于产生大麻素糖苷的方法,包括在允许糖基转移酶将核苷酸糖苷的糖基部分转移至大麻素的条件下,使大麻素受体与条目19至72所述的一种或多种大麻素糖基转移酶和条目15至18所述的一种或多种核苷酸糖苷接触。
139.一种产生大麻素的方法,包括根据条目129至136所述的方法产生大麻素糖苷,以及使大麻素糖苷经受一个或多个去糖基化步骤。
140.根据条目139所述的方法,其中去糖基化可以通过将大麻素糖苷与一种或多种酶孵育来实现,该酶选自葡糖苷酶、果胶酶、阿拉伯糖酶、纤维素酶、葡聚糖酶、半纤维素酶和木聚糖酶。
141.根据条目140所述的方法,其中一种或多种酶选自β-葡糖苷酶、β-β葡聚糖酶、果胶裂解酶、果胶酶和多半乳糖醛酸酶。
142.根据条目139至141所述的方法,其中去糖基化步骤在体外进行。
143.一种发酵液,包括条目128所述的细胞培养物中包括的大麻素糖苷。
144.根据条目143所述的发酵液,其中至少50%(诸如至少75%、诸如至少95%、诸如至少99%)的基因修饰的宿主细胞被分解。
145.根据条目143至144所述的发酵液,其中至少50%(诸如至少75%、诸如至少95%、诸如至少99%)的固体细胞材料已经与液体分离。
146.根据条目144至145所述的发酵液,进一步包括选自以下的一种或多种化合物:
a)产生所述大麻素糖苷的起作用的生物合成代谢途径的前体或产物;
b)包括微量金属、维生素、盐、酵母氮源基础、YNB和/或氨基酸的补充营养物;并且
其中所述大麻素糖苷的浓度为至少1mg/l液体。
147.一种大麻素糖苷,包括与选自以下的糖共价连接的大麻素苷元或大麻素糖苷:木糖;鼠李糖;半乳糖;N-乙酰葡糖胺;N-乙酰半乳糖胺;和阿拉伯糖。
148.根据条目147所述的大麻素糖苷,其中所述大麻素糖苷选自大麻素 -1'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖苷;大麻素-1'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖苷;大麻素 -1'-O-β-D-N-乙酰半乳糖胺;大麻素-1'-O-β-D-纤维二糖苷;大麻素-1'-O-β-D- 龙胆二糖苷;大麻素-1'-O-β-D-木糖基-3'-O-β-D-木糖苷;大麻素-1'-O-α-L- 鼠李糖基-3'-O-β-D-鼠李糖苷;大麻素-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;大麻素 -1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;和大麻素-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺。
149.根据条目148所述的基因修饰的宿主细胞,其中,该大麻素糖苷选自CBD-1'-O-β-D-纤维二糖苷;CBD-1'-O-β-D-龙胆二糖苷;CBD-1'-O-β-D- 木糖基-3'-O-β-D-木糖苷CBD-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷;CBD-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBD-1'-O-β-D-N-乙酰葡糖胺 -3'-O-β-D-N-乙酰氨基葡糖苷;CBD-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;CBD-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺;CBDV-1'-O-β-D-纤维二糖苷;CBDV-1'-O-β-D-龙胆二糖苷;CBDV-1'-O-β-D- 木糖基-3'-O-β-D-木糖苷;CBDV-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷;CBDV-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBDV-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;CBDV-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;CBDV-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺; CBG-1'-O-β-D-纤维二糖苷;CBG-1'-O-β-D-龙胆二糖苷;CBG-1'-O-β-D-木糖基-3'-O-β-D-木糖苷CBG-1'-O-α-L-鼠李糖基-3'-O-α-L-鼠李糖苷; CBG-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;CBG-1'-O-β-D-N-乙酰葡糖胺 -3'-O-β-D-N-乙酰氨基葡糖苷;CBG-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;CBG-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺; THC-1'-O-β-D-纤维二糖苷;THC-1'-O-β-D-龙胆二糖苷;THC-1'-O-β-D-木糖苷;THC-1'-O-α-L-鼠李糖苷;THC-1'-O-β-D-半乳糖苷;THC-1'-O-β-D-N-乙酰氨基葡糖苷;THC-1'-O-β-D-阿拉伯糖苷;THC-1'-O-β-D-N-乙酰氨基半乳糖苷;CBN-1'-O-β-D-纤维二糖苷;CBN-1'-O-β-D-龙胆二糖苷; CBN-1'-O-β-D-木糖苷;CBN-1'-O-α-L-鼠李糖苷;CBN-1'-O-β-D-半乳糖苷; CBN-1'-O-β-D-N-乙酰氨基葡糖苷;CBN-1'-O-β-D-阿拉伯糖苷; CBN-1'-O-β-D-N-乙酰氨基半乳糖苷;CBDA-1'-O-β-D-纤维二糖苷; CBDA-1'-O-β-D-龙胆二糖苷;CBDA-1'-O-β-D-木糖苷;CBDA-1'-O-α-L-鼠李糖苷;CBDA-1'-O-β-D-半乳糖苷;CBDA-1'-O-β-D-N-乙酰氨基葡糖苷; CBDA-1'-O-β-D-阿拉伯糖苷;CBDA-1'-O-β-D-N-乙酰氨基半乳糖苷; CBC-1'-O-β-D-纤维二糖苷;CBC-1'-O-β-D-龙胆二糖苷;CBC-1'-O-β-D-木糖苷;CBC-1'-O-α-L-鼠李糖苷;CBC-1'-O-β-D-半乳糖苷;CBC-1'-O-β-D-N- 乙酰氨基葡糖苷;CBC-1'-O-β-D-阿拉伯糖苷;和CBC-1'-O-β-D-N-乙酰氨基半乳糖苷。
150.一种大麻素糖苷,包括通过1,4-或1,6-糖苷键与糖基部分共价连接的大麻素苷元或大麻素糖苷。
151.根据条目148所述的大麻素糖苷,其中大麻素糖苷选自 CBD-1'-O-β-D-龙胆二糖苷和CBD-1'-O-β-D-纤维二糖苷。
152.一种组合物,包括条目143至146所述的发酵液和/或条目147至 151所述的大麻素糖苷以及一种或多种剂、添加剂和/或赋形剂。
153.根据条目152所述的组合物,其中发酵液以及一种或多种剂、添加剂和/或赋形剂呈干燥固体形式。
154.根据条目152所述的组合物,其中发酵液以及一种或多种剂、添加剂和/或辅料呈液体稳定形式。
155.根据条目154所述的组合物,其中将组合物精制成适合人或动物摄取的饮料,并且其中与未糖基化的大麻素相比,大麻素糖苷的水溶解度增加。
156.根据条目153所述的组合物,其中,将组合物精制成适合人或动物摄取的食品,并且其中与未糖基化的大麻素相比,大麻素糖苷水溶解度增加。
157.一种用于制备药物制剂的方法,包括将条目147至151所述的大麻素糖苷或其前药或条目152至156所述的组合物与一种或多种药物级赋形剂、添加剂和/或佐剂混合。
158.根据条目157所述的方法,其中该药物制剂是粉末、片剂、胶囊、硬咀嚼剂和/或软锭剂或口香糖的形式。
159.根据条目157所述的方法,其中该药物制剂是液体药物溶液的形式。
160.一种可获自条目157至159所述的方法的药物制剂。
161.一种可获自条目157至159所述的方法的药物制剂,用作药物或前药用途。
162.根据条目161所述的制剂,用于治疗哺乳动物中选自以下的疾病: NASH、癫痫、呕吐、恶心、癌症、多发性硬化症、痉挛、慢性疼痛、厌食症、食欲不振、帕金森病、德拉韦综合征(婴儿严重肌阵挛癫痫)、伦诺克斯-加斯托综合征、物质(药物)滥用、糖尿病、癫痫发作、恐慌症、社交焦虑症(SAD)、广泛性焦虑症(GAD)、焦虑症、广场恐惧症、婴儿痉挛症(韦斯特综合征)、银屑病、疱疹后神经痛、运动神经元疾病、肌萎缩侧索硬化、图雷特综合征、抽动障碍、大脑性瘫痪、移植物抗宿主病(GVHD)、克罗恩病 (区域性肠炎)、炎症性肠病、脆性X综合征、双相情感障碍(躁狂抑郁症)、骨关节炎、亨廷顿病、精神分裂症、自闭症、不安腿综合征、人类免疫缺陷病毒(HIV)感染(AIDS)、高血压、肝纤维化、肝损伤、普拉德-威利综合征(PWS)、创伤后应激障碍(PTSD)、脂肪肝、青光眼、炎症性疾病、艰难梭菌感染、结直肠肿瘤、炎症性肠病、肠病、肠易激综合征、溃疡性结肠炎、认知障碍、脑缺氧、纤维化、睡眠呼吸暂停、运动神经元病、抗菌素耐药性、细菌感染和COVID-19感染。
163.一种用于治疗哺乳动物中疾病的方法,包括向所述哺乳动物给药治疗有效量的条目160所述的药物制剂或条目147至151所述的大麻素糖苷。
164.根据条目163所述的方法,其中,疾病选自NASH、癫痫、呕吐、恶心、癌症、多发性硬化症、痉挛、慢性疼痛、厌食症、食欲不振、帕金森病、德拉韦综合征(婴儿严重肌阵挛癫痫)、伦诺克斯-加斯托综合征、物质(药物)滥用、糖尿病、癫痫发作、恐慌症、社交焦虑症(SAD)、广泛性焦虑症(GAD)、焦虑症、广场恐惧症、婴儿痉挛症(韦斯特综合征)、银屑病、疱疹后神经痛、运动神经元疾病、肌萎缩侧索硬化、图雷特综合征、抽动障碍、大脑性瘫痪、移植物抗宿主病(GVHD)、克罗恩病(区域性肠炎)、炎症性肠病、脆性X综合征、双相情感障碍(躁狂抑郁症)、骨关节炎、亨廷顿病、精神分裂症、自闭症、不安腿综合征、人类免疫缺陷病毒(HIV)感染 (AIDS)、高血压、肝纤维化、肝损伤、普拉德-威利综合征(PWS)、创伤后应激障碍(PTSD)、脂肪肝、青光眼、炎症性疾病、艰难梭菌感染、结直肠肿瘤、炎症性肠病、肠病、肠易激综合征、溃疡性结肠炎、认知障碍、脑缺氧、纤维化、睡眠呼吸暂停、运动神经元病、抗菌素耐药性、细菌感染和COVID-19感染。
参考文献
Gajewski,J.,Pavlovic,R.,Fischer,M.,Boles,E.和Grininger,M.(2017). 用于短链脂肪酸生产的工程真菌从头脂肪酸合成,自然通讯,8, 1–8(Gajewski,J.,Pavlovic,R.,Fischer,M.,Boles,E.,&Grininger, M.(2017).Engineering fungal de novo fattyacid synthesis for short chain fatty acid production.Nature Communications,8,1–8.)https://doi.org/10.1038/ncomms14650
Gietz,R.D.和Woods,R.A.(2002).通过乙酸锂/单链运载体DNA/聚乙二醇法转化酵母.酶学方法,350(2001),87-96.(Gietz,R.D.,&Woods,R. A.(2002).Transformationof yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycolmethod.Methods in Enzymology,350(2001), 87–96.)https://doi.org/10.1016/S0076- 6879(02)50957-5
Grote,A.,Hiller,K.,Scheer,M.,Münch,R.,
Figure BDA0003490979370001251
B.,Hempel,D. C.和Jahn,D.(2005).JCat:一种使靶基因的密码子使用适应其潜在表达宿主的新工具.核酸研究,33(SUPPL.2),526–531(Grote,A.,Hiller,K.,Scheer,M., Münch,R.,
Figure BDA0003490979370001252
B.,Hempel,D.C.,&Jahn,D.(2005).JCat:A novel tool to adapt codon usage of a targetgene to its potential expression host.Nucleic Acids Research,33(SUPPL.2),526–531.)https://doi.org/10.1093/nar/gki376
Gueldener,U.,Heinisch,J.,Koehler,G.J.,Voss,D.和Hegemann,J. H.(2002).用于芽殖酵母中Cre介导的多基因敲除的第二组loxP标记盒.核酸研究,30(6),e23(Gueldener,U.,Heinisch,J.,Koehler,G.J.,Voss,D.,& Hegemann,J.H.(2002).A secondset of loxP marker cassettes for Cre-mediated multiple gene knockouts inbudding yeast.Nucleic Acids Research,30(6),e23)。获自http:// www.ncbi.nlm.nih.gov/pubmed/11884642%0Ahttp://www.pubmedcentral.nih.gov/ articlerender.fcgiΔartid=PMC101367
Jensen,N.B.,Strucko,T.,Kildegaard,K.R.,David,F.,Maury,J., Mortensen,U.H.,……Borodina,I.(2014).EasyClone:酿酒酵母中多个基因的迭代染色体整合方法,FEMS酵母研究,14(2),238-248(Jensen,N.B., Strucko,T.,Kildegaard,K.R.,David,F.,Maury,J.,Mortensen,U.H.,… Borodina,I.(2014).EasyClone:Method for iterativechromosomal integration of multiple genes in Saccharomyces cerevisiae.FEMSYeast Research,14(2), 238–248)。https://doi.org/10.1111/1567-1364.12118
Jessop-Fabre,M.M.,
Figure BDA0003490979370001261
T.,Stovicek,V.,Dai,Z.,Jensen,M.K.,Keasling,J.D.和Borodina,I.(2016).EasyClone-MarkerFree:通过 CRISPR-Cas9将基因无标记整合至酿酒酵母中的载体工具包,生物技术杂志,11(8),1110-1117(Jessop-Fabre,M.M.,
Figure BDA0003490979370001262
T.,Stovicek,V.,Dai,Z., Jensen,M.K.,Keasling,J.D.,&Borodina,I.(2016).EasyClone-MarkerFree:A vector toolkit for marker-less integration ofgenes into Saccharomyces cerevisiae via CRISPR-Cas9.Biotechnology Journal,11(8),1110–1117)。 https://doi.org/10.1002/biot.201600147
van Rossum,H.M.,Kozak,B.U.,Pronk,J.T.和van Maris,A.J.A.(2016). 工程酿酒酵母中的细胞溶质乙酰-辅酶A供应:途径化学计量、自由能守恒和氧化还原辅助因子平衡,代谢工程,36,99-115(van Rossum,H.M.,Kozak, B.U.,Pronk,J.T.,&van Maris,A.J.A.(2016).Engineering cytosolic acetyl-coenzyme A supply in Saccharomycescerevisiae:Pathway stoichiometry, free-energy conservation and redox-cofactorbalancing.Metabolic Engineering, 36,99–115)。https://doi.org/10.1016/ j.ymben.2016.03.006
Shi,S.,Chen,Y.和Siewers,V.(2014).提高丙二酰辅酶A衍生代谢物的产量,微生物学(MBio),5(3),e01130-14(Shi,S.,Chen,Y.,&Siewers,V. (2014).ImprovingProduction of Malonyl Coenzyme A-Derived Metabolites. MBio,5(3),e01130-14)。https://doi.org/10.1128/mBio.01130-14
Luo,X.,Reiter,M.A.,d’Espaux,L.,Wong,J.,Denby,C.M.,Lechner, A.,……Keasling,J.D.(2019).在酵母中完全生物合成大麻素及其非天然类似物,自然2019,1(Luo,X.,Reiter,M.A.,d’Espaux,L.,Wong,J.,Denby,C.M., Lechner,A.,…Keasling,J.D.(2019).Complete biosynthesis of cannabinoids and their unnaturalanalogues in yeast.Nature 2019,1)。 https://doi.org/10.1038/s41586-019-0978-9
Degenhardt,F.,Stehle,F.和Kayser,O.(2017).大麻素的生物合成,大麻及相关病理学手册:生物学、药理学、诊断和治疗,爱思唯尔公司(Degenhardt, F.,Stehle,F.,&Kayser,O.(2017).The Biosynthesis of Cannabinoids. Handbook of Cannabis andRelated Pathologies:Biology,Pharmacology, Diagnosis,and Treatment.ElsevierInc.)。https://doi.org/10.1016/B978-0-12-800756-3.00002-8
Mackenzie,P.I.,Owens,I.S.,Burchell,B.等人(1997)UDP糖基转移酶基因超家族:基于进化分歧的建议命名更新,药物遗传学,7,255-269 (Mackenzie,P.I.,Owens,I.S.,Burchell,B.et al.(1997)The UDP glycosyltransferase gene superfamily:recommended nomenclature update based on evolutionarydivergence.Pharmacogenetics,7,255–269)。
实施例
实施例
材料和方法
材料
在本文的实施例中使用的例如用于缓冲液和底物的化学品是至少试剂级的商业产品。
菌株
BY4723是常见的酿酒酵母菌株,源自S288C,并且可获自例如美国典型培养物保藏中心(ATCC#200885)。
BY4741是常见的酿酒酵母菌株,源自S288C,并且可获自例如 Euroscarf(Y00000)。
BL21(DE3)是常见的大肠杆菌菌株,可获自例如新英格兰生物实验室 (NewEngland Biolabs)(C2527I)。
DH5α是常见的大肠杆菌菌株,可获自例如赛默飞世尔科技 (ThermoFisherScientific)(18265017)。
XJb(DE3)自溶菌株是常见的大肠杆菌菌株,可获自例如Zymo Research(T3051)。
用于从实施例2、4、7、14-15和21的培养基中提取和回收大麻素的方法:
部分I.
在培养酿酒酵母或大肠杆菌后,如下从培养基中提取大麻素或大麻素糖苷。样品最初用2U/OD溶细胞酶(Zymo Research)处理(2h,30℃,800 rpm)(大肠杆菌培养跳过该步骤),然后用乙酸乙酯/甲酸(0.05%(v/v))以2:1 的比率提取和打珠(bead-beating)(30s-1,3min)。然后将样品以12,000g离心1min,并丢弃无机级分。然后重复用乙酸乙酯/甲酸提取。然后将剩余的有机级分在真空箱中在50℃下蒸发至干,然后将干燥的提取物重悬于乙腈 /H2O/甲酸(80%/20%/0.05%(v/v/v))中。最后,使用Ultrafree-MC色谱柱(0.22 μm孔径,聚偏二氟乙烯(PVDF)膜)过滤样品。
部分II.
替代地,大肠杆菌或酿酒酵母中的大麻素或大麻素糖苷的全细胞肉汤提取如下。将细胞培养物与100%甲醇1:1混合,添加玻璃珠,并使用打珠机(例如FastPrep)使细胞爆裂打开。将样品以12,000g离心1min,并且上清液直接用于分析。
用于实施例2、4、7-14、16-18和20-21的分析程序:
部分I.
HPLC分析在配备DAD检测器的Agilent Technologies 1100系列上进行。在Kinetex 2.6μm XB-C18色谱柱(100×2.1mm,2.6μm,
Figure BDA0003490979370001281
飞诺美(Phenomenex))上实现分离。溶剂:H2O中的0.05%(v/v)三氟乙酸和MeCN 中的0.05%(v/v)三氟乙酸分别作为流动相A和B。梯度条件:0.0-23min 1%-99%B;23.1-25.0min 99-1%和25.1-27.0min 2%B。流动相流量为400 μL/min。柱温保持在30℃。在230和254nm处获得UV光谱。自动进样器温度设置为10℃±2℃。使用可靠的参比标准品鉴定大麻素。使用用大麻素标准溶液的一系列浓度绘制的标准校准曲线进行定量。
部分II.
LC-MS分析通过UPLC耦连到电喷雾离子源(ESI)(沃特世(Waters),米尔福德(Milford),马萨诸塞州(MA)的三重四极杆质谱仪进行。将1μL提取的样品进样至LC-MS系统,并使用配备C18 BEH(1.7μm,2.1x50mm)预装柱(沃特世,米尔福德,马萨诸塞州)的C18BEH(1.7μm)柱在反相中实现分离,并且流动相由
Figure BDA0003490979370001291
级水中的0.1%甲酸(西格玛-奥德里奇)(A)和MS 级乙腈中的0.1%甲酸的(B)组成,流速为0.6mL/min。Masslynx软件(1.6版) 用于仪器控制,而Markerlynx用于数据集成。使用1.0min内从50%B至 100%B的线性梯度实现大麻素分离,并保持0.5min,然后在下次进样前将色谱柱在50%B下重新平衡0.7min。该方法的总运行时间为2.2min。质谱仪使用多反应监测(MRM)模式在负离子模式下运行。使用的两个最丰富的跃迁是357.12>178.99和357.12>245.06。两次跃迁的锥电压均设置为54 V,而第一次跃迁的碰撞能量设置为22eV,以及第二次跃迁的碰撞能量设置为28eV。SIM模式用于检测。对于所有不同的MS分析,毛细管电压设置为2.2kV。为了定量,在可能的情况下,在甲醇中制备单独的1mg/mL 的大麻素储备溶液。随后,在甲醇:水(1:1,v/v)中制备工作溶液,以获得 (0.16-20)μM的浓度范围。大麻素糖苷最初在非靶向方法中鉴定,并且后来在SIM模式下使用每个糖苷分子的m/z预测值进行半定量。
部分III.
替代地,为了更好地分离具有多种糖的亲水性大麻素糖苷, LC-MS/Q-TOF分析在Dionex UltiMate 3000四元快速分离UHPLC+聚焦系统(赛默飞世尔科技,盖默灵(Germering),德国(Germany))联用Compact microTOF-Q质谱仪(布鲁克(Bruker),不来梅港市(Bremen),德国)上进行。在Kinetex 1.7μm XB-C18色谱柱(150×2.1mm,1.7μm,
Figure BDA0003490979370001292
飞诺美)上实现分离。溶剂:H2O中的0.05%(v/v)甲酸和MeCN分别作为流动相A和B。梯度条件:梯度(A):0.0-2.0min 2%B;2.0-.0-25.0min 2-100%B, 25.0-27.5min100%B,27.5-28.0min 100-2%B以及28.0-30.0min 2%B。梯度(B):0.0-1.0min 10%B;1.0-24.0min 10-85%B;24.0-25.0min 85-100%B,25.0-27.5min 100%B,27.5-28.0min100-2%B以及28.0-30.0 min 2%B。流动相流量为300μL/min。柱温保持在30℃下。在220、230、 240和280nm处获得UV光谱。Compact micrOTOF-Q质谱仪(布鲁克,不来梅港市,德国)配备以正离子模式运行的电喷雾离子源。离子喷雾电压保持在4500V,干气体温度为250℃。氮气用作干气体(8L/min)、雾化气体(2.5 bar)和碰撞气体。碰撞能量设置为10eV。MS和MS/MS光谱在50至1000 amu的m/z范围内以2Hz的采样率采集。甲酸钠簇用于质量校准。
在实施例8、13、16、18和20的体外酶测定中提取和回收大麻素和糖基化的大麻素:
部分I.
通过将整个反应混合物在100%甲醇中稀释4倍,同时从体外酶测定中提取疏水性大麻素和亲水性大麻素糖苷。对于LC-MS/Q-TOF分析,样品在 50%MeOH中进一步稀释10倍,并如上所述进行分析。
部分II.
替代地,亲水性大麻素糖苷从体外糖基化测定中提取,并与疏水性大麻素底物分离,如下。乙酸乙酯提取使用反应混合物以1:1的比率进行。有机和水性级分通过重力分离并分别收集。分离出的水性级分用乙酸乙酯1:1 再提取2次。如上所述通过HPLC分析有机相和水性相两者的一小部分以确证大麻素糖苷的存在。使用旋转蒸发器蒸发含有大麻素糖苷的相。将所得干级分重悬于100%甲醇中并超声处理5分钟。通过以1:4(v/v)的比率添加冰冷100%丙酮并在-20℃下孵育过夜来沉淀重悬液中的蛋白质。通过在8000rpm下离心30min去除蛋白质沉淀物并回收上清液。在冷冻干燥回收的上清液之前重复离心以蒸发甲醇和丙酮。在上样至制备型HPLC进行纯化之前,将所得干燥沉淀物重悬于20%DMSO中。在配备有DAD检测器的Agilent 1200制备型HPLC上纯化大麻素糖苷。在
Figure BDA0003490979370001301
5μm C18(2)LC 柱(150×21.2mm,5μm,
Figure BDA0003490979370001302
飞诺美)上实现分离。溶剂:H2O中的0.01%(v/v) 三氟乙酸和MeCN中的0.01%(v/v)三氟乙酸分别作为流动相A和B。梯度条件:0-1min 5%B;1-5min 5-40%B;5-20min 40-80%B;20-21min 80-100%B;21-24min 100%B;24-25min 100-5%B.流动相流量为15 mL/min。柱温为室温。在220、230和280nm处获得UV光谱。级分收集器在5-20min内每0.5min收集一次级分,取决于大麻素糖苷。收集包含基于230nm处的UV光谱峰的级分,并通过HPLC(如上所述)分析亚级分以确证身份,并冷冻干燥至干燥以回收作为粉末的纯化大麻素糖苷。如上所述,通过LC-MS/QTOF分析纯化化合物的精确质量。
实施例1-用于产生大麻素的基因修饰的酿酒酵母菌株的构建
部分I.
基于Gajewski,Pavlovic,Fischer,Boles和Grininger,自然通讯;DOI: 10.1038/ncomms14650,2017(Gajewski,Pavlovic,Fischer,Boles,&Grininger, Nature Comm;DOI:10.1038/ncomms14650,2017.)所述的工作进行产生己酸的酿酒酵母菌株的构建。替代地,可以使用WO2016156548的程序。
www.yeastgenome.org的酵母基因组数据库(SGD)中公开的PDR12基因缺失通过如下方式实现。用引物将LoxP侧翼SpHis5盒从pUG27扩增 (Gueldener等人,2002),其中引物与PDR12的上游和下游区具有60bp的添加的同源性。在含有20g/L葡萄糖减去组氨酸补充剂(SC-His)的合成培养基上的转化和选择产生PDR12缺失的菌株。
使用由(Jessop-Fabre等人,2016)描述的EasyClone无标记系统使用核酸内切酶(诸如MAD7)实现从大麻素生物合成途径的基因的整合(https://www.inscripta.com/)。如下表(表1-3)中所述构建靶向基因组中预定位置的整合质粒。构建这些质粒的质粒骨架获自Addgene (https://www.addgene.org/)。根据(Gietz&Woods,2002),通过用NotI(新英格兰生物实验室公司(New England Bio Labs Inc.))限制性消化使质粒线性化,并与靶向每个基因组位置的gRNA质粒一起转化至酿酒酵母中。将转化体接种于选择性培养基上。
表1.用于构建产生大麻素的酿酒酵母菌株的整合质粒
Figure BDA0003490979370001311
Figure BDA0003490979370001321
表2.用于构建整合质粒的生物块
Figure BDA0003490979370001322
Figure BDA0003490979370001331
表3.用于扩增生物块的引物
Figure BDA0003490979370001332
Figure BDA0003490979370001341
Figure BDA0003490979370001351
使用JCAT算法(Grote等人,2005)对所有异源基因进行密码子优化以在酿酒酵母中表达,由GeneArt合成并置于强酿酒酵母组成型启动子和终止子的控制下。使用PhusionU聚合酶(ThermoScientific)进行生物块的扩增。
部分II.
替代地,可以如下构建产生大麻素的菌株。可以如上所述构建产生己酸的菌株,或者替代地可以将己酸外源添加到培养基中。大麻素生物合成途径的基因使用定制的过表达质粒整合至预定的基因组“着陆区(landing pad)”中,类似于(Mikkelsen等人,2012)所述。线性整合片段通过NotI消化定制设计的质粒产生,该质粒含有强组成型酿酒酵母启动子和终止子,并且两侧是上游和下游同源区,以促进同源重组的组装。为了便于在单个基因组位点组装多个整合质粒,设计了上游和下游同源臂,使得在NotI消化 (New England BioLabs Inc.)后,线性整合片段可以重组为单个线性整合片段并整合至靶基因组位点。为了选择已成功整合感兴趣的片段的转化体,可以如上所述使用核酸内切酶(诸如MAD7),或替代地可以将选择标记(诸如 LEU2)并入线性整合片段并转化到本领域已知的亮氨酸营养缺陷型酿酒酵母菌株。为了减少假阳性的发生,可以将选择标记分为2个线性整合片段,诸如Rec 1和Rec 2,使得仅在Rec 1和Rec 2整合片段成功同源重组后才能生成功能性LEU2选择标记,如图1所示。
基因经过密码子优化以在酵母中表达,并由Twist Biosciences合成并克隆到定制的整合质粒中(表4)。根据(Gietz&Woods,2002),用NotI(新英格兰生物实验室公司)限制性消化线性化后,将质粒转化到酿酒酵母中。将转化体接种于选择性培养基上。
表4.用于构建产生大麻素的酿酒酵母菌株的整合质粒
Figure BDA0003490979370001361
Figure BDA0003490979370001371
实施例2-在基因修饰的酿酒酵母菌株中产生大麻素
部分I.
酵母菌株在500μL液体合成完全培养基(SC)或具有20g/L葡萄糖减尿嘧啶补充剂(SC-Ura)的合成完全培养基中在30℃、300rpm下在具有透气密封的2mL微量滴定板中预培养24h。随后,将50μL酵母预培养物转移至 450μL SC或SC-Ura中,其中含有20g/L实时补料(FIT)基本培养基 (Enpresso),其中含有0.3%酶,或其他合适的碳源,诸如20g/L葡萄糖并在 30℃、300rpm下生长72h。细胞在含有己酸(1mM)、丁酸(1mM)、大麻素生物合成途径的其他中间体,或未添加补充剂(如上所述从头产生脂肪酸的菌株)的培养基中孵育。孵育后,如上所述提取和分析大麻素。如上所述,所有分析均使用HPLC或LC-MS,并在可能的情况下使用可靠的分析标准品。由于生物合成生产产生大麻素的酸形式,而脱羧形式通常是生物活性形式,在一些方面,脱羧大麻素通过将蒸发的大麻素提取物在110℃下加热 50分钟进行制备,然后重悬于<乙腈/H2O/甲酸(80%/20%/0.05%(v/v/v))中。在一些方面,如上所述,在进一步提取之前,通过将细胞培养液在80℃下直接加热50分钟来制备脱羧大麻素。
部分II.
替代地,将酵母菌株在不含氨基酸补充剂的合成培养基中在30℃和 300rpm下按需要预培养过夜,以维持对引入的表达质粒和/或整合盒的选择。随后将10μL细胞培养物转移至490μL的减氨基酸补充剂的合成培养基中,根据需要补充有20g/L葡萄糖、20g/L乙醇、1mM己酸或1mM丁酸大麻素生物合成路径的其他中间体(或其组合)。细胞在30℃和300rpm下孵育3天,如前所述提取和分析大麻素。脱羧大麻素通过以下制备:将蒸发的大麻素提取物在110℃下加热50分钟,然后重悬于乙腈/H2O/甲酸 (80%/20%/0.05%(v/v/v))中。在一些方面,如上所述,在进一步提取之前,通过将细胞培养液在80℃下直接加热50分钟来制备脱羧大麻素。
实施例3-用于产生大麻素的基因修饰的大肠杆菌菌株的构建
如下将大麻素生物合成途径引入大肠杆菌。使用添加限制性消化位点的引物从合成DNA扩增基因,并将其克隆至pETDuet-1、pETACYCDuet-1 和pCDFDuet-1双表达载体(Novagen)中。将质粒转化至大肠杆菌菌株 BL21(DE3)并分别在氨苄青霉素、氯霉素和链霉素上筛选出成功的转化体。所使用的质粒(表5)、生物块(表6)和引物(表7)的概要如下所示。
表5.构建用于在大肠杆菌中设计大麻素生物合成的质粒
Figure BDA0003490979370001381
表6.用于构建质粒的生物块
Figure BDA0003490979370001391
表7用于扩增生物块的引物。
Figure BDA0003490979370001392
Figure BDA0003490979370001401
Figure BDA0003490979370001411
实施例4-在基因修饰的大肠杆菌菌株中产生大麻素
大肠杆菌菌株在500μL补充有氨苄青霉素、氯霉素和链霉素 (LB+AmpChlorStrep)的液体LB培养基中在37℃、300rpm的具有透气密封的2mL微量滴定板中预培养24h。随后将50μL预培养物转移至450μl 的LB+AmpChlorStrep中,并添加20g/L葡萄糖,并在37℃、300rpm下培养24h。细胞在含有己酸(1mM)、丁酸(1mM)、大麻素生物合成途径的其他中间体或未添加脂肪酸补充剂(如上所述从头产生脂肪酸的菌株)的培养基中进一步孵育,并添加多肽表达诱导剂。孵育后,如上所述提取和分析大麻素。如上所述,所有分析均使用LC-MS或HPLC,并在可能的情况下使用可靠的分析标准品。由于生物合成生产产生大麻素的酸形式,而脱羧形式通常是生物活性形式,在一些方面,脱羧大麻素通过将蒸发的大麻素提取物在110℃下加热50分钟进行制备,然后重悬于<乙腈/H2O/甲酸 (80%/20%/0.05%(v/v/v))中。在一些方面,如上所述,在进一步提取之前,通过将细胞培养液在80℃下直接加热50分钟来制备脱羧大麻素。
实施例5-用于产生大麻素糖苷的酿酒酵母菌株的构建
部分I.
在酿酒酵母中表达的基因由GeneArt进行密码子优化和合成。使用添加U2 USER克隆位点的引物对基因进行PCR扩增,并使用强组成型启动子和终止子,使用(Jensen等人,2014)所述的EasyClone系统将基因克隆至组成型表达载体pCfB132中。在没有尿嘧啶的情况下,通过在培养基上铺板来选择转化体。所使用的质粒(表8)、生物块(表9)和引物(表10)的概述如下。质粒骨架可获自Addgene(https://www.addgene.org/)
表8.构建用于在酿酒酵母中过表达糖基转移酶的质粒
Figure BDA0003490979370001421
Figure BDA0003490979370001431
Figure BDA0003490979370001441
表9.构建酿酒酵母中糖基转移酶质粒的生物块。
Figure BDA0003490979370001442
Figure BDA0003490979370001451
表10.用于构建生物块的引物
Figure BDA0003490979370001452
Figure BDA0003490979370001461
Figure BDA0003490979370001471
Figure BDA0003490979370001481
Figure BDA0003490979370001491
部分II.
替代地,在酿酒酵母中表达的基因经过密码子优化、由Twist Biosciences 合成并克隆至质粒中。基因被克隆至酵母着丝粒表达载体p413TEF中,该载体含有TEF1强组成型启动子、CYC1终止子和HIS3营养缺陷型标记。 p413TEF质粒骨架可获自ATCC(ATCC#87362)。在没有组氨酸的情况下,通过在培养基上铺板来选择转化体。质粒的概要如下表11所述。
表11.构建用于在酿酒酵母中过表达糖基转移酶的质粒
Figure BDA0003490979370001492
Figure BDA0003490979370001501
Figure BDA0003490979370001511
实施例6-构建用于产生大麻素糖苷的大肠杆菌菌株
部分I.
在大肠杆菌中表达的糖基转移酶基因由GeneArt合成。使用添加限制性位点的引物对基因进行PCR扩增,并使用标准限制性/连接克隆将基因克隆至pRSFDuet-1表达质粒中。通过在含有卡那霉素的培养基上铺板来选择转化体。将质粒转化至DH5α、“Arcticexpress”(安捷伦科技(Agilent technologies))或Xjb-autolysis BL21(Zymo research)大肠杆菌菌株或先前实施例构建的大肠杆菌菌株中。使用的质粒(表12)、生物块(表13)和质粒(表 14)概述如下
表12.构建用于将糖基转移酶引入大肠杆菌的质粒。
Figure BDA0003490979370001521
Figure BDA0003490979370001531
表13.用于在大肠杆菌中构建糖基转移酶质粒的生物块
Figure BDA0003490979370001532
Figure BDA0003490979370001541
表14.用于构建生物块的引物。
Figure BDA0003490979370001551
Figure BDA0003490979370001561
Figure BDA0003490979370001571
Figure BDA0003490979370001581
Figure BDA0003490979370001591
部分II.
替代地,在大肠杆菌中表达的糖基转移酶基因针对大肠杆菌表达进行了密码子优化,并通过使用SpeI/XhoI限制性位点的标准限制性连接由Twist Bioscience合成并克隆至定制的质粒载体(pRSGLY,由GeneArt合成)中。该定制的载体包含LacI操纵子、AmpR盒、复制起点和多克隆位点,两侧是T7启动子和终止子。此外,5’端还含有核酶结合位点(RBS)和6xHis标签用于后续的蛋白质纯化。将完全组装的质粒转化至大肠杆菌DH5α菌株或大肠杆菌XJb(DE3)自溶菌株(Zymo Research)中。使用的质粒如表15所示。
表15.构建用于在大肠杆菌中表达糖基转移酶的质粒
Figure BDA0003490979370001601
Figure BDA0003490979370001611
Figure BDA0003490979370001621
实施例7-在基因修饰的菌株中产生大麻素化合物
部分I.
大麻素糖苷在大肠杆菌或酿酒酵母菌株中通过补料葡萄糖(从头产生)、脂肪酸(例如己酸和丁酸)、大麻素生物合成途径中的其他中间体(例如橄榄酸、divarinolicacid、大麻萜酚酸)、最终的大麻素本身(生物转化)或其组合产生。大肠杆菌细胞在Lysogeny肉汤中与适当的抗生素一起孵育,加入多肽表达诱导剂,在30℃下持续振荡72h。酿酒酵母细胞在具有所需氨基酸补充剂以补偿营养缺陷型的合成培养基中孵育,并在30℃下持续振荡72h。如上所述提取和分析大麻素和大麻素糖苷。如有需要,将UDP-糖底物添加至生长培养基中。替代地,将催化糖转化为活化糖(例如蔗糖转化为UDP- 葡萄糖)的酶和/或催化活化糖相互转化(例如UDP-葡萄糖转化为UDP-鼠李糖)的酶引入基因修饰的菌株。
部分II.
替代地,可以使用细胞内源性UDP糖池(例如,由酿酒酵母和大肠杆菌天然产生的UDP葡萄糖)。
实施例8-糖基化大麻素受体中糖基转移酶性能的体外检测
对于糖基转移酶性能的体外研究,构建以表达糖基转移酶的大肠杆菌菌株的粗裂解物通过以下制备:将菌株放入含有1mL的含卡那霉素的 NZCYM细菌培养液的无菌96深孔板中。将样品在37℃下孵育过夜,以 200rpm振荡。第二天,将50μl的每种培养物转移到新的无菌96深孔板中,其含有1mL含卡那霉素和多肽表达诱导剂的NZCYM细菌培养液。样品在 20℃下孵育,以200rpm振荡20h。此后,将板在4℃下以4000rpm离心 10min。倾倒出上清液后,将50μl的包含Tris-HCl、MgCI2、CaCI2和蛋白酶抑制剂的缓冲液加入每个孔中,并通过在4℃下以200rpm振荡5min来重悬细胞。然后将每个孔的内容物(即细胞浆液)转移到PCR板并在-80℃下冷冻过夜。冷冻细胞浆在室温下融化最长达30min。如果融化混合物由于细胞裂解而不粘稠,则将样品再次冻融。当样品几乎融化时,将25μl的包含DNase和MgCI2的结合缓冲液加入到每个孔中。PCR板在室温下孵育5 min,以500rpm振荡,直到样品变得不那么粘稠。最后,样品以4000rpm 离心5min,并且上清液用于将大麻素转化为其糖基化衍生物。根据表16在体外进行转化。碱性磷酸酶由新英格兰生物实验室(M0371S)提供。大麻素受体溶解在DMSO中。
表16.用于体外测量糖基转移酶活性的反应装置。
Figure BDA0003490979370001631
Figure BDA0003490979370001641
将反应混合物在30℃下孵育过夜。通过加入30μl 100%DMSO终止反应。所得混合物用90μl 50%DMSO进一步稀释用于LC-MS分析和表现最佳的糖基转移酶的分级。
替代地,以下实施例13的方案用于该体外检测。
实施例9-糖基化大麻素的水溶解度检测
部分I.
按照生产商的说明,使用用于溶解度测定的
Figure BDA0003490979370001642
HTS-PCF过滤板(Merck)确定水溶解度。将纯化的大麻素糖苷溶解在DMSO中至初始浓度为20mM。如上所述,使用LC-MS/QTOF确定溶液中大麻素糖苷的定量。
部分II.
替代地,可以通过在LC-MS/QTOF分析过程中测量化合物的保留时间来对水性溶解度进行定性测量。由于极性化合物在运行期间将在较早的保留时间洗脱,而且由于极性是水性溶解度的直接指标,因此可以进行比较评估。水性溶解度的定性测量还可以通过计算分子的分配系数(cLogP)来进行。cLogP是衡量溶质在水部分与有机部分溶解多少的量度,cLogP较低的分子比cLogP较高的分子更能溶于水。可以使用化合物的分子结构和专用软件计算cLogP。ChemSketch(ACD Labs)用于计算大麻素和大麻素糖苷的 cLogP。
如上所述,通过LC-MS/QTOF分析一系列大麻素葡糖苷,并测量保留时间(RT)并与其LogP(cLogP)计算值进行比较。如下表17所示,大麻素糖苷具有的保留时间比大麻素短,表明它们更易溶于水。此外,大麻素二糖苷的具有的保留时间比单糖苷短,并且大麻素三糖苷的保留时间比二糖苷短,总体上表明向大麻素中添加糖基团导致水溶解度连续增加。测量的保留时间也与LogP计算值相关。
表17.QTOF分析期间的保留时间(RT)以及大麻素和大麻素糖苷的LogP 计算值
Figure BDA0003490979370001651
Figure BDA0003490979370001661
部分III.
替代地,水性溶解度通过如下热力学溶解度测定法进行测定。在玻璃小瓶中称量2.5mg检测化合物,加入0.5mL磷酸盐缓冲盐水(pH=7.4),并短暂涡旋样品。然后将样品在小瓶滚轴系统上在室温下孵育过夜,以使尽可能多的化合物溶于溶液中。孵育之后,将水溶液一式两份过滤(0.45μM孔径),滤液用100%甲醇1:1稀释。必要时进一步稀释样品并通过HPLC分析。通过与由可靠的分析标准品制成的标准曲线进行比较来确定溶液中化合物的浓度。
如上所述测量CBD和CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB6)的水性热力学溶解度,并确定它们溶解度的定量测量。如下表18所示,OB6具有比CBD显著更高的水溶解度,在室温下在PBS(pH=7.4)中达到11.4±0.75 mM的溶解度。CBD的溶解度低于HPLC机器的检测限,通过稀释可靠的分析CBD标准,发现检测限为0.5μM,表明CBD的最大溶解度为0.5μM。
表18.CBD和CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB6)在室温下在 PBS缓冲液pH7.4中的热力学溶解度(以mM计)。BDL:低于检测限。数据表示为重复实验的平均值和标准偏差。
Figure BDA0003490979370001671
实施例10-糖基化大麻素的化学稳定性检测
部分I.
通过在DMSO中制备10mM储备溶液,以及然后在甘氨酸缓冲液(pH 8–11)、PBS(pH7–8)和乙酸盐缓冲液(pH 4-6)中稀释至5μM,确定大麻素糖苷的化学稳定性。溶液在37℃下孵育,以0、60、120、180、240和300 分钟的间隔取样。如上所述使用LC-MS分析所有样品。
部分II.
替代地,大麻素糖苷的化学稳定性在碱性、酸性、氧化和热应激下测定如下。在100%甲醇中制备25mM大麻素和大麻素糖苷的储备溶液。将 15μL与5μL 400mM HCl溶液(最终pH=1.1)、400mM NaOH溶液(最终 pH=12.5)、12%12%H2O2溶液(最终浓度3%)或H2O pH7.0混合。将酸性、碱性和氧化性样品在30℃下孵育24h,而水中的样品在80℃下孵育24h。还制备了环境条件下的对照,其中将15μL的大麻素或大麻素糖苷添加到5 μL H2O pH 7.0中,并在30℃下孵育。24h后,将样品置于冰上,并向每个样品中加入60μL冰冷的100%甲醇。将样品离心并转移到HPLC小瓶中进行分析。通过与可靠的分析标准品进行比较,对大麻素或大麻素糖苷的剩余浓度进行定量。通过与可靠的分析标准品进行比较来确定降解产物的存在。
CBD、CBD-1'-O-β-D-glucoside(OB1)-葡糖苷(OB1)和CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB6)暴露于上述氧化、碱性、酸性和热条件下,并通过HPLC分析通过测量在给定条件下暴露24h后残留在溶液中的化合物的量对其降解进行定量,并表示为相对于环境条件下的对照在24h暴露后剩余的百分比(%)。还测量了已知CBD降解产物THC的累积,以暴露24h 后累积的百分比表示。如表19所示,CBD在所有检测条件下均不稳定,并且特别是在酸性和碱性条件下降解为THC。CBD在碱性条件下特别不稳定,暴露24h后仅残留2.26%。相反,在所有检测条件下,特别是在100%残留的碱性条件下,暴露24h后,OB1和OB6的含量显著更高。而少量的 THC-1'-O-β-D-葡糖苷(OB20)在酸性条件下检测到OB1,而暴露于任何条件下的OB6样品均未检测到THC或THC-葡糖苷。同样重要的是,在任何条件下均未检测到OB1和OB6的CBD苷元,从而表明葡糖苷键在极端条件下具有稳定性。
表19.CBD、CBD-1'-O-β-D-葡糖苷(OB1)和CBD-1'-O-β-D-葡糖基 -3'-O-β-D-葡糖苷(OB6)在酸性、碱性、氧化和热应激条件下的化学稳定性。将底物在每种条件下孵育24h,然后通过HPLC进行分析。显示的是溶液中残留底物的%和已知降解产物THC(和THC-1'-O-β-D-葡糖苷(OB20))相对于对照(在30℃下无应激孵育的底物,pH7.0)的累积%。每个测定中使用的底物以粗体表示。数据显示为生物重复的平均值。ND;未检测到,NA;不适用。
Figure BDA0003490979370001681
Figure BDA0003490979370001691
实施例11-糖基化大麻素的血浆稳定性检测
大麻素糖苷的血浆稳定性通过以下确定:在37℃下在人血浆(西格玛) 中孵育1μM并以0、60、120、180、240和300分钟的时间间隔取样。如上所述使用LC-MS分析所有样品。维拉帕米和丙胺太林用作高稳定性和低稳定性参比品。
实施例12-糖基化大麻素的肝微粒体稳定性检测
部分I.
大麻素糖苷的肝微粒体稳定性通过将2μM分子与补充有NADPH的 HepaRGTM人肝微粒体(西格玛)在37℃下孵育来确定。以0、5、15、30、 45和60分钟的间隔取样并如上所述进行分析。维拉帕米(快速清除)和地西泮(低清除)用作参比品。
部分II.
替代地,如下测定大麻素糖苷的肝微粒体稳定性。HepaRGTM混合的人肝微粒体(西格玛)(最终蛋白质浓度=0.5mg/mL)与丙甲菌素(25μg/mg)、0.1 M磷酸盐缓冲液(pH=7.4)和检测化合物(DMSO中的最终浓度为1μM)混合并在37℃下孵育,然后加入NADPH(终浓度1mM)和UDP-葡糖醛酸(终浓度1mM)以启动反应。将化合物孵育0、5、15、30和45分钟,以及然后通过加入比率为1:3(v/v)的乙腈终止反应。将反应物在4℃下以3000rpm离心20min以沉淀蛋白质。蛋白质沉淀后,将内标添加到样品上清液中,并通过LC-MS分析以测量每个时间点残留的化合物浓度,通过与可靠的分析标准品进行比较来实现定量。
如上所述,对CBD、CBD-1'-O-β-D-葡糖苷(OB1)和CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB6)进行体外肝微粒体稳定性并测定每种化合物的固有清除率(CLin)和半衰期(t1/2)。如下表20所示,发现虽然OB1具有比CBD 更低的肝微粒体稳定性(由更高的内在清除率和更短的半衰期表示),但OB6 具有显著更高的肝微粒体稳定性,如半衰期增加50倍和内在清除率相应减少50倍所示。
表20.CBD、CBD-1'-O-β-D-葡糖苷(OB1)和CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB6)的肝微粒体稳定性。显示的是每种化合物的固有清除率(CLint)和半衰期(t1/2)。数据表示为不同时间点(0、5、15、40、45min)的5 个生物重复的平均值和标准偏差。
Figure BDA0003490979370001701
Figure BDA0003490979370001711
实施例13-糖基化大麻素中糖基转移酶性能的体外检测
对于糖基化大麻素中糖基转移酶性能的体外研究,纯化的糖基转移酶制备如下:
用表达感兴趣的糖基转移酶的大肠杆菌XJb(DE3)菌株接种5mL的2x 浓缩LB培养基+氨苄青霉素(50μg/mL),并在30℃下振荡孵育过夜。第二天,将细胞培养物转移到500mL的2x浓缩LB培养基+氨苄青霉素(50 μg/mL)中,并在30℃下振荡孵育过夜。第二天,将细胞培养物转移到1L 的2x浓缩LB培养基+氨苄青霉素(50μg/mL)+3mM阿拉伯糖+0.1mM IPTG中。细胞在20℃下振荡孵育24h。第二天,通过在4℃下以46500xg 离心10min来收集细胞。将细胞重悬于20mL冰冷的GT缓冲液(50mM Tris-HCl pH7.4+1mM苯甲磺酰氟+1cOmpleteTM,微型,无EDTA蛋白酶抑制剂Cocktail片(罗氏(Roche))中。将重悬的物质转移到50mL falcon管中并在-80℃下保持至少15min。然后将Falcon管在室温下融化,随着试管融化,添加以下试剂;溶解在MilliQ水中的2.6mM MgCl2,1mM CaCl2, 250μL的1.4mg/ml溶液(西格玛)。轻轻倒转试管混合,然后在37℃下孵育5min。然后将结合缓冲液加入管中(50mM Tris-HCl pH7.4、10mM咪唑、 500mM NaCl,11.25mL MilliQ水)并用HCl调节pH值至7.4。将混合物在 4℃下以15550xg离心15min,将上清液转移到新鲜的50mL falcon管中并再次在4℃下以48400xg离心20min以去除任何剩余的细胞碎片。在酶制剂离心的同时,将3mL HIS-Select(可获自西格玛P6611)柱材料添加到新的 50mL管中,并通过添加最高达50mL的MilliQ水进行洗涤,以2000xg离心2min并弃去上清液。重复该洗涤步骤。最后,将MilliQ水添加至HIS-Select材料中至约50%的体积。通过Miracloth(可获自默克Millipore) 将来自离心酶制剂的收集的上清液转移到含有HIS-Select材料的管中,以及然后在4℃下通过倒转轻轻振荡孵育2h。2h后,将混合物在4℃下以 2000xg离心4分钟,弃去上清液。剩余的HIS-Select材料用1x结合缓冲液 (50mM Tris-HCl、0.5M NaCl、10mM咪唑,pH 7.4)洗涤两次,并在4℃下以2000xg离心4分钟。将HIS-Select材料重悬于5mL 1x结合缓冲液中并转移至
Figure BDA0003490979370001721
色谱柱(可获自BioRad,7311550)。HIS-Select材料保持在4℃,并用1x结合缓冲液冲洗两次,装柱并使其滴入。最后,通过添加 7.5mL洗脱缓冲液(50mM Tris-HCl,500mM咪唑,pH7.4)以及收集流出液来从HIS-Select材料中洗脱纯化的糖基转移酶。酶可立即用于体外酶测定或在-20℃下储存在50%甘油中待用。
根据表21进行各种大麻素至大麻素糖苷的体外转化。碱性磷酸酶由新英格兰生物实验室(M0371S)提供。将大麻素溶解在甲醇中。UDP-糖(例如 UDP-葡萄糖)由商业供应商(例如西格玛)提供或通过从市售UDP-糖体外酶促转化产生,如实施例21所示。
表21.用各种大麻素在体外测量糖基转移酶活性的反应设置(setup)。
Figure BDA0003490979370001722
Figure BDA0003490979370001731
根据需要放大或缩小反应混合物。将反应混合物在30℃下在不振荡的情况下孵育24小时。提取和分析如上文针对本实施例所述进行。为了确认所产生的大麻素糖苷的身份,如上所述使用LC-MS/QTOF来确认每个检测到的分子的预期质量和碎片模式。通过将大麻素底物和大麻素糖苷的峰面积与可靠的分析标准品(如果可用)进行比较,对大麻素糖苷的产量进行定量,在没有底物的情况下,通过与大麻素苷元的可靠分析标准品进行比较来实现定量。通过测量培养24h后底物的减少和产物的增加,计算由特定糖基转移酶将底物转化为大麻素糖苷的百分比。总体上,使用UDP-葡萄糖、 UDP-鼠李糖、UDP-木糖、UDP-半乳糖、UDP-葡糖醛酸和UDP-N-乙酰葡糖胺对大麻素CBD、CBDV、CBDA、THC、CBN、CBG和11-nor-9-羧基 -THC的大麻素糖基化进行检测。
为该筛选中产生的每种大麻素糖苷提供了相应的结构ID,每个分子的结构如图4所示。生成的LC-MS/QTOF色谱图的实例如图5所示。
使用CBD作为大麻素受体产生的大麻素糖苷。
发现一系列糖基转移酶可以催化CBD转化为一系列不同的CBD糖苷。表22示出了产生的所有CBD-糖苷和示例性糖基转移酶,其以相应的转化%催化每个反应。
表22.由糖基转移酶体外产生的CBD-葡糖苷
Figure BDA0003490979370001732
Figure BDA0003490979370001741
Figure BDA0003490979370001751
表23进一步示出了每种化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了每种CBD-糖苷的结构。
表23.糖基转移酶体外产生的每种CBD糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001761
Figure BDA0003490979370001771
对于若干种CBD-糖苷,发现多种糖基转移酶可以以不同的转化效率催化反应。表24-30示出了产生CBD-糖苷的糖基转移酶以及转化%效率。
表24.催化CBD转化为OB1(CBD→CBD-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001781
表25.催化CBD转化为OB13(CBD→CBD-1'-O-α-L-鼠李糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001782
Figure BDA0003490979370001791
表26.催化CBD转化为OB9(CBD→CBD-1'-O-β-D-木糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001792
表27.催化CBD转化为OB6(CBD→CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001801
表28.催化CBD转化为OB10(CBD→CBD-1'-O-β-D-木糖基-3'-O-β-D- 木糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001802
表29.催化CBD转化为OB7(CBD→CBD-1'-O-β-D-三-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001811
表30.催化CBD转化为OB8(CBD→CBD-1'-O-β-D-葡糖基-3'-O-β-D-二-葡糖基)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001812
使用CBDV作为大麻素受体产生的大麻素糖苷。
发现一系列糖基转移酶可以催化CBDV转化为一系列不同的CBDV糖苷。表31示出了产生的所有CBDV-糖苷和示例性糖基转移酶,其以相应的转化%催化每个反应。
表31.由糖基转移酶体外产生的CBDV-葡糖苷
Figure BDA0003490979370001813
Figure BDA0003490979370001821
表32进一步示出了每种化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了每种CBDV-糖苷的结构。
表32.糖基转移酶体外产生的每种CBDV-糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001831
对于若干种CBDV-糖苷,发现多种糖基转移酶可以以不同的转化效率催化反应。表33-34提供了被表明可以产生CBDV-糖苷的糖基转移酶以及转化效率%的列表。
表33.催化CBDV转化为OB24(CBDV→CBDV-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001841
表34.催化CBDV转化为OB25(CBDV→(CBDV→CBDV-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001851
使用CBDA作为底物产生的大麻素糖苷。
发现一系列糖基转移酶可以催化CBDA转化为OB31。表35示出了产生的CBDA-糖苷和示例性糖基转移酶,其以相应的转化%催化每个反应。
表35.糖基转移酶体外产生的CBDA-葡糖苷
Figure BDA0003490979370001852
表36进一步示出了化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了CBDA- 糖苷的结构。
表36.糖基转移酶体外产生的CBDV-糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001861
发现多种糖基转移酶可以以不同的转化效率催化该反应。表37提供了被表明可以产生CBDA-糖苷的糖基转移酶以及转化效率%的列表。
表37.催化CBDA转化为OB31(CBDA→CBDA-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001862
Figure BDA0003490979370001871
使用CBG作为底物产生的大麻素糖苷。
发现一系列糖基转移酶催化CBG转化为一系列不同的CBG糖苷。表 38示出了产生的所有CBG-糖苷和示例性糖基转移酶,其以相应的转化%催化每个反应。
表38.由糖基转移酶体外产生的CBG-葡糖苷。
Figure BDA0003490979370001872
Figure BDA0003490979370001881
表39进一步示出了每种化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了每种CBG-糖苷的结构。
表39.糖基转移酶体外产生的每种CBG-糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001882
Figure BDA0003490979370001891
Figure BDA0003490979370001901
对于若干种CBG-糖苷,发现多种糖基转移酶可以以不同的转化效率催化反应。表40-41提供了被表明可以产生CBG-糖苷的糖基转移酶以及转化效率%的列表。
表40.催化CBG转化为OB32(CBG→CBG-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001902
Figure BDA0003490979370001911
表41.催化CBG转化为OB33((CBG→CBG-1’-O-β-D-葡糖基-3'-O-β-D- 葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001912
Figure BDA0003490979370001921
使用THC作为底物产生的大麻素糖苷。
发现一系列糖基转移酶催化THC转化为一系列不同的THC糖苷。表 42示出了产生的所有THC-糖苷和示例性糖基转移酶,其以相应的转化%催化每个反应。
表42.由糖基转移酶体外产生的THC-葡糖苷。
Figure BDA0003490979370001922
表43进一步示出了每种化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了每种THC-糖苷的结构。
表43.糖基转移酶体外产生的每种THC-糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001923
Figure BDA0003490979370001931
对于OB20,发现多种糖基转移酶可以以不同的转化效率催化反应。表 44提供了被表明可以产生THC-糖苷的糖基转移酶以及转化效率%的列表。
表44.催化THC转化为OB20(THC→THC-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001932
使用CBN作为底物产生的大麻素糖苷。
发现一系列糖基转移酶催化CBN转化为至少一种CBN-糖苷。表45示出了产生的所有CBN-糖苷和示例性酶,其以相应的转化%催化每个反应。
表45.由糖基转移酶体外产生的CBN-葡糖苷。
Figure BDA0003490979370001941
表46进一步示出了每种化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了每种CBN-糖苷的结构。
表46.糖基转移酶体外产生的每种CBN-糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001942
对于OB23,发现多种糖基转移酶可以以不同的转化效率催化反应。表 47提供了被表明可以产生CBN-糖苷的糖基转移酶以及转化效率%的列表。
表47.催化CBN转化为OB23(CBN→CBN-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001943
Figure BDA0003490979370001951
使用11-nor-9-羧基-THC作为底物产生的大麻素糖苷。
发现一系列糖基转移酶催化11-nor-9-羧基-THC转化为一系列11-nor-9- 羧基-THC-糖苷。表48示出了产生的所有11-nor-9-羧基-THC-糖苷和示例性糖基转移酶,其以相应的转化%催化每个反应。
表48.糖基转移酶体外产生的11-nor-9-羧基-THC-葡糖苷。
Figure BDA0003490979370001952
Figure BDA0003490979370001961
表49进一步示出了每种化合物的保留时间(RT)、LogP计算值(clogP)、预期和测量质量以及通过LC-MS/QTOF分析确定的碎裂模式,从而证实了每种11-nor-9-羧基-THC-糖苷(OB41、42)的结构。
表49.糖基转移酶体外产生的每种11-nor-9-羧基-THC-糖苷的保留时间、cLogP、预期和测量质量以及碎裂模式。
Figure BDA0003490979370001962
对于OB41,发现多种糖基转移酶可以以不同的转化效率催化反应。表 50提供了被表明可以产生11-nor-9-羧基-THC-糖苷的糖基转移酶以及转化效率%的列表。
表50.催化11-nor-9-羧基-THC转化为OB41(11-nor-9-羧基 -THC→11-nor-9-羧基-THC-1'-O-β-D-葡糖苷)的糖基转移酶及计算的转化效率。ND:未检测到。
Figure BDA0003490979370001971
Figure BDA0003490979370001981
进一步发现,一系列糖基转移酶可以使用大麻素作为糖受体,从而产生相当多的新的大麻素糖苷。在筛选中,发现了可以催化各种不同且高特异性反应的酶。发现糖基转移酶可以特异性地产生单糖苷(例如由Pt88G (SEQ ID NO:147、148)产生的CBD-1'-O-β-D-葡糖苷(OB1))、二糖苷(例如由Cp73B(SEQ ID NO:191、192))产生的CBD-1'-O-β-D-葡糖基-3'-O-β-D- 葡糖苷(OB6))、三糖苷(例如由At73C5(SEQ ID NO:107、108)产生的 CBG-1'-O-β-D-葡糖基-3'-O-β-D-二-葡糖苷(OB33))以及甚至四糖苷(例如由 Cs73Y(SEQ ID NO:157、158)产生的CBG-1'-O-β-D-四-木糖苷(OB40))。
还发现一系列糖基转移酶可以利用一系列不同的UDP-糖,例如发现 Cs73Y(SEQID NO:157、158)可以利用UDP-葡萄糖、UDP-木糖、UDP- 鼠李糖、UDP-葡糖醛酸、UDP-半乳糖和UDP-N-乙酰葡糖胺,并将这些糖附连接至各种大麻素。
根据计算的转化%,发现许多糖基转移酶具有高活性,能够以极高效率催化大麻素糖苷的产生。若干种酶在24h内将100%的大麻素苷元转化为相应的大麻素糖苷(例如由Cp73B(SEQ ID NO:191、192)产生的 CBN-1'-O-β-D-二-葡糖苷(OB23)和由Pt78G(SEQ IDNO:165、166))产生的 CBG-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB33))。
还发现大量的酶可以催化产生大麻素糖苷。该体外筛选总共鉴定了51 种酶。
此外,还检测了从甜叶菊中分离的并针对在现有技术中描述的能够糖基化一系列大麻素在大肠杆菌中的表达进行密码子优化的糖基转移酶 Sr76G1(SEQ ID NO:123、124)对一系列大麻素和大麻素糖苷底物的糖基转移酶活性。然而发现Sr76G1(SEQ ID NO:123、124)可以使葡萄糖附连至大麻素葡糖苷的葡萄糖部分(例如将CBD-1'-O-β-D-葡糖苷(OB1)转化为 CBD-1'-O-β-D-昆布二糖苷(OB2))。然而出乎意料的是,使用任何大麻素苷元作为底物未检测到糖基转移酶活性。
实施例14-大麻素底物在大肠杆菌中体内生物转化为糖基化衍生物
为了证明大麻素在体内转化为大麻素糖苷,根据实施例6部分II构建了含有糖基转移酶表达质粒PL-5(At73C5_GA)(SEQ ID NO:107、108)、 PL-182(Ha88B_2_GA)(SEQ IDNO:149、150)和PL-214(Cs73Y_GA)(SEQ ID NO:157、158)的大肠杆菌菌株,产生大肠杆菌菌株EC-5、EC-182和 EC-214。还包括Sr76G1表达质粒(PL-55(Sr76G1_GA(SEQ ID NO:123,124)) (产生大肠杆菌菌株EC-55)以检测在体外观察到的活性缺失是否也在体内观察到。随后将菌株在37℃下在10mL预培养管中的补充有氨苄青霉素的 5mL的LB培养基中孵育过夜。随后,将细胞接种到96深孔板中补充有氨苄青霉素的500μL的LB培养基中,起始OD600为0.1,并在30℃下孵育 6小时。然后将大麻素底物溶解在乙醇中,并与合适的诱导剂(IPTG)一起添加到培养基中,终浓度如下:
乙醇:20g/L
大麻素底物:250μM
IPTG:0.15mM
将细胞与添加的乙醇、大麻素底物和IPTG再培养66小时。如上所述,通过HPLC分析提取和分析大麻素糖苷。对大麻素浓度的降低和大麻素糖苷的累积进行定量,并计算每种糖苷的转化百分比。如下表51所示,表达糖基转移酶的大肠杆菌菌株可以将一系列大麻素转化为其相应的糖苷。
表51.通过表达糖基转移酶的大肠杆菌菌株将大麻素体内生物转化为大麻素糖苷。示出了大麻素转化到大麻素糖苷的转化率。ND;未检测到,WT 对照;XJb(DE3)亲本菌株。
Figure BDA0003490979370001991
Figure BDA0003490979370002001
结果表明选定的糖基转移酶可以在体内产生一系列大麻素糖苷,结果还证实在体外观察到的Sr76G1(SEQ ID NO:123、124)活性缺乏在体内被复制。如体外测定中所见的,一些糖基转移酶可以非常高效的产生大麻素糖苷,例如Cs73Y(SEQ ID NO:157、158)将进料的CBN100%转化为OB23。此外,结果表明,在大肠杆菌中表达的糖基转移酶可以利用细胞内源性 UDP-葡糖库进行反应,而无需另外补充这种底物。使用THC和11-nor-9- 羧基-THC作为底物未检测到活性,但是在体外检测到活性,表明大肠杆菌将大麻素转化为大麻素糖苷的能力可能有限。
实施例15-在酿酒酵母中大麻素底物向糖基化衍生物的体内生物转化
前面的实施例已表明纯化的糖基转移酶可以在体外将一系列底物转化为大麻素糖苷,以及在大肠杆菌中表达的糖基转移酶通过在培养基中补料大麻素底物并使用细胞内源性供应UDP-葡萄糖也能如此。为了证明在酿酒酵母体内大麻素向大麻素糖苷的生物转化,将先前显示在大肠杆菌中体外和体内催化一系列大麻素转化为大麻素糖苷的糖基转移酶Cs73Y(SEQ ID NO:207、208)针对在酿酒酵母中的表达进行密码子优化,克隆到着丝粒表达载体p413TEF(产生质粒PL-388(p413TEF:Cs73Y)并转化至酿酒酵母菌株 BY4741中(产生菌株SC-1)。将SC-1在30℃下在含有20g/L葡萄糖的 SC-His培养基中预培养过夜,然后将10μL细胞培养物转移到490μL的含有20g/L葡萄糖并补充有溶解在100%乙醇中的各种大麻素的SC-His培养基中并在30℃下孵育3天。培养基中大麻素的终浓度为250μM并且乙醇的终浓度为20g/L。如上所述制备和分析样品。如表52所示,表达糖基转移酶Cs73Y的SC-1可以将一系列大麻素高效地转化为它们各自的单糖苷、二糖苷和三糖苷。
表52.通过表达糖基转移酶Cs73Y的酿酒酵母菌株SC-1将大麻素体内生物转化为大麻素糖苷。示出了大麻素转化到大麻素糖苷的转化率。ND;未检测到,WT对照;BY4741亲本菌株。
Figure BDA0003490979370002011
发现,SC-1可以将所有检测的大麻素转化为大麻素糖苷,而且效率非常高。对于除THC和11-nor-9-羧基-THC之外的所有检测大麻素,发现SC-1 将所有添加的大麻素转化为大麻素糖苷。此外,虽然在表达糖基转移酶的大肠杆菌培养物中未检测到THC和11-nor-9-羧基-THC糖苷的产生,但在酿酒酵母培养物中检测到THC和11-nor-9-羧基-THC糖苷。这不仅表明大麻素成功导入细胞并且细胞内源性UDP-葡萄糖供应足以进行反应,还表明与大肠杆菌相比,酿酒酵母是产生大麻素糖苷的更优的宿主。
实施例16-糖基化大麻素的肠道渗透性检测
大麻素和糖基化大麻素的肠道渗透性通过测量跨Caco-2细胞膜的双向转运进行确定。Caco-2细胞被用作人肠道上皮的体外模型并用于评估潜在药物的肠道渗透性。将检测化合物添加到Caco-2细胞汇合单层的顶端或基底外侧,并通过使用LC-MS/QTOF监测出现单层另一侧的检测化合物以测量渗透性。在进行双向检测时,流出率(ER)由B-A和A-B渗透率的比率计算得出。从ATCC获得的Caco-2细胞用于40-60代传代。将细胞以1×105 个细胞/cm2接种到Millipore Multiscreen Transwell板上。将细胞在DMEM 中培养,每两至三天更换一次培养基。在第20天进行渗透性研究。细胞培养和测定孵育在37℃、5%CO2和95%相对湿度的气氛中进行。在测定当天,通过在加热至37℃的期望pH值下用Hanks平衡盐溶液(HBSS)冲洗顶端和基底外侧表面两次来制备单层。然后将细胞与HBSS在顶端和基底外侧隔室中在期望pH值下孵育40min以稳定生理参数。在DMSO中制备10 mM大麻素和大麻素糖苷溶液,然后用测定缓冲液稀释,得到10μM的检测化合物终浓度(DMSO终浓度为1%v/v)。荧光完整性标记荧光黄也包括在溶液中。分析标准品由检测化合物DMSO稀释液制备并转移到缓冲液中,保持1%v/v DMSO浓度。为了评估A-B渗透性,从顶端隔室中取出HBSS 并替换为测试化合物溶液。然后将顶端隔室插入物放入含有新鲜缓冲液(含有1%v/v DMSO)的配套板中。为了评估B-A渗透性,从配套板上取出HBSS 并替换为检测化合物溶液。将新鲜缓冲液(含1%v/v DMSO)添加到顶端隔室插入物中,然后将其放入配套板中。在120min时,将顶端隔室插入物和配套板分开,顶端和基底外侧样品被稀释以供分析。一式两份评估检测化合物渗透性。已知渗透性特征的化合物在每个测定板上作为对照运行。检测和对照化合物通过如上所述的LC-MS/QTOF进行定量。起始浓度(C0)由溶液确定,并且实验回收率由C0以及顶端和基底外侧隔室浓度两者计算得出。整个实验过程中单层的完整性通过使用荧光分析监测荧光黄渗透进行检查。每种化合物的渗透系数(Papp)通过以下方程计算Papp=(dQ/dt)/(C0× A),其中dQ/dt是药物穿过细胞的渗透率,C0是零时的供体隔室浓度,并且A是细胞单层的面积。C0获自配量溶液的分析。流出率(ER)根据A-B和 B-A数据的平均Papp值计算。这获自:ER=Papp(B-A)/Papp(A-B)。回收率%由以下方程计算;回收率%=(实验结束时供体和受体隔室中的总化合物)/(存在的初始化合物)×100。
测量A至B和B至A两方向的平均渗透系数(Papp)、平均底物回收率以及CBD、CBD-1'-O-β-D-葡糖苷(OB1)和CBD-1'-O-β-D-葡糖基-3'-O-β-D- 葡糖苷(OB6)的相应流出率。CBD糖苷是使用糖基转移酶并如上所述纯化。如下表53所示,与未修饰的CBD相比,OB1在两个方向上具有显著更高的渗透系数和更高的外排比,总体上表明肠道渗透性和流出改善。对于OB6,虽然渗透系数较低,但产生的流出率高于CBD和OB1,表明分子从肠道的流出有所改善。此外,结果清楚地示出,糖基化提高了回收率,在 OB1和OB6的两个隔室中观察到的回收率逐渐提高。Caco-2渗透性测定中化合物的低回收率可能表明存在溶解度差、化合物与板结合、Caco-2细胞代谢或化合物在细胞单层中累积的问题。
表53.Caco-2双向渗透性测定中CBD、CBD-1'-O-β-D-葡糖苷(OB1)和 CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷(OB6)肠道渗透性的体外测量。结果计算为重复实验的平均值和标准偏差。方向A→B;自顶端至基底外侧隔室扩散,方向B→A;自基底外侧至顶端隔室扩散。Papp;渗透系数
Figure BDA0003490979370002031
Figure BDA0003490979370002041
实施例17-在酿酒酵母中从头产生糖基化大麻素
为了证明大麻素糖苷的从头产生,如前所述,将用于产生CBDA的异源生物合成途径引入酿酒酵母野生型菌株BY4741,产生菌株SC-CBDA。此外,示出糖基化在质粒PL-388(p413TEF:Cs73Y)上表达的一系列大麻素的糖基转移酶Cs73Y(SEQ ID NO:207、208)被转移到该菌株中,产生菌株 SC-CBDAGLY。用于构建这些菌株的质粒示于表54,并且引入的所得生物合成途径示于图3。
表54.用于构建产生SC-CBDA和SC-CBDAGLY大麻素的酿酒酵母菌株的质粒。
Figure BDA0003490979370002042
Figure BDA0003490979370002051
随后如先前所述在添加20g/L葡萄糖和1mM己酸减去亮氨酸和组氨酸补充剂(SC-Ura+His)的合成培养基中培养菌株,并如前所述制备和分析样品。如下表55所示,引入大麻素生物合成途径(SC-CBDA)导致产生1.97μM CBDA,进一步引入糖基转移酶Cs73Y导致产生2.03μM CBDA-1'-O-β-D- 葡糖苷(OB31)。如上所述加热细胞培养液导致从SC-CBDA细胞培养物中产生0.87μM CBD和从SC-CBDAGLY细胞培养物中产生1.54μM CBD-1'-O-β-D-葡糖苷(OB1)。
表55.在工程化的酿酒酵母菌株中从头产生大麻素和大麻素糖苷。ND;未检测到。数据以μM表示,并表示为重复实验的平均值。细胞在补充有 20g/L葡萄糖和1mM己酸的SC-Ura+His培养基中培养3天。
CBDA OB31 CBD OB1
SC-CBDA 1.97 ND 0.87 ND
SC-CBDAGLY ND 2.03 ND 1.54
实施例18-从蔗糖和大麻素底物产生大麻素糖苷的体外酶级联反应
在前面的实施例中,体外糖基转移酶分析需要添加“活化”糖(例如UDP- 葡萄糖),这通常是非常昂贵的试剂,此外,其他活化糖(例如UDP-鼠李糖) 无法商购获得,且必须定制合成,其成本高且难度大。在体内,虽然酿酒酵母和大肠杆菌能够天然产生UDP-葡萄糖,但它们的产量很低,而且不能产生其他活化糖,从而限制了它们在体内产生多种大麻素糖苷的适用性。为了促进大麻素糖苷的低成本生产,不仅使用葡萄糖,而且使用替代糖,建立了酶促级联反应以将大麻素和单糖蔗糖转化为各种大麻素糖苷。级联分为3个步骤,在步骤1中,蔗糖和尿苷二磷酸(UDP)通过GmSuSy(SEQ ID NO:209、210)转化为UDP-葡萄糖,另外生成果糖作为副产物。在步骤2 中,使用一系列酶将UDP-葡萄糖相互转化为替代的UDP-糖。例如,通过 BsGalE将UDP-葡萄糖转化为UDP-半乳糖,多种酶也可以用于通过其他 UDP-糖中间体产生UDP-糖。例如,通过AtUGDH1将UDP-葡萄糖转化为 UDP-葡糖醛酸,组合通过AtUXS3将UDP-葡糖醛酸转化为UDP-木糖。在步骤3中,糖基转移酶将活化糖和大麻素受体转化为相应的大麻素糖苷。例如,通过Cs73Y(SEQ ID NO:157、158)将UDP-鼠李糖和CBD转化为 CBD-1'-O-β-D-鼠李糖苷(OB13)。可以相互转化UDP-糖的酶的实例如下表 (表56)所示。
表56.用于UDP-糖相互转化的酶。
Figure BDA0003490979370002061
替代地,为了产生UDP-鼠李糖,不是使用全长AtRHM2基因(SEQ ID NO:219、220),为了更好的表达和更高的活性,AtRHM2可以分为N-和 C-末端结构域AtRHM2-N(SEQ ID NO:217、218)和AtRHM2-C(SEQ ID NO: 215、216)分别催化脱水、以及差向异构化和还原。替代地,可以将所有三种(全长AtRHM2(覆盖氨基酸1-667)、AtRHM2-N(覆盖氨基酸1-370)和AtRHM2-C(覆盖氨基酸371-667))混合以增加UDP-鼠李糖的产生。
级联反应可以在单个反应中进行,替代地,也可以将步骤1、2和3拆分为不同的反应并根据需要进行组合。
使用纯化的GmSuSy和Cs73Y酶与UDP-糖互变酶和所需辅助因子的不同组合,用CBD体外证明了这种用于产生大麻素糖苷的酶级联反应。酶被纯化并如实施例13中所述进行体外测定,并且反应混合物如表57中所示设置。根据每个单独反应的需要添加酶和辅助因子。如上所述提取和分析样品。
表57.用于体外产生具有替代糖的大麻素糖苷的反应设置。
Figure BDA0003490979370002071
如下表58所示,通过添加不同的酶组合,可以从蔗糖和CBD高效产生各种CBD-二-葡糖苷。
表58.通过添加糖转化酶的不同组合,将CBD和蔗糖转化为各种CBD 糖苷。ND;未检测到
Figure BDA0003490979370002081
实施例19-使用糖基转移酶产生新的分子
本发明的糖基转移酶已经揭示并使得生产一系列迄今为止未知的大麻素糖苷成为可能,这些大麻素糖苷可以大致分为以下几类:
表59.由本发明的酶产生的新的大麻素糖苷的类别。还显示了每个类别的示例性分子以及可用于产生该分子的相应一种或多种酶和SEQ ID NO。
Figure BDA0003490979370002082
Figure BDA0003490979370002091
本发明的酶可以用于产生以下分子:
表60.由本发明的酶产生的新的大麻素糖苷的列表。还显示了可以用于产生每个分子的酶和相应的SEQ ID NO。
Figure BDA0003490979370002092
Figure BDA0003490979370002101
Figure BDA0003490979370002111
Figure BDA0003490979370002121
Figure BDA0003490979370002131
Figure BDA0003490979370002141
Figure BDA0003490979370002151
Figure BDA0003490979370002161
Figure BDA0003490979370002171
Figure BDA0003490979370002181
Figure BDA0003490979370002191
Figure BDA0003490979370002201
Figure BDA0003490979370002211
Figure BDA0003490979370002221
Figure BDA0003490979370002231
Figure BDA0003490979370002241
Figure BDA0003490979370002251
Figure BDA0003490979370002261
实施例20-结合多种糖基转移酶催化大麻素底物转化为具有交替的糖- 糖键的大麻素糖苷
本文所述的糖基转移酶可以广泛地分为对大麻素苷元有活性的糖基转移酶或对大麻素糖苷有活性的糖基转移酶。后一组不是将糖部分附连至大麻素分子上的游离羟基上,而是将糖部分附连至大麻素糖苷的糖基团上。在实施例13中,发现了一系列仅对大麻素苷元有活性的糖基转移酶(例如 PL-159(Pt88G_GA)(SEQ ID NO:147、148))以及一系列对大麻素苷元和大麻素糖苷两者均具有活性的糖基转移酶。例如,发现PL-214(Cs73Y_GA)(SEQ ID NO:157、158)产生一系列多糖大麻素糖苷,包括大麻素键上的糖以及糖键上的糖。在实施例13中,还发现一些糖基转移酶仅对大麻素糖苷有活性,并且在糖糖基化反应中特异性地催化糖。这些酶中的两种 (PL-55(Sr76G1_GA)(SEQ ID NO:123、124)和PL-32(OsEUGT11_GA)(SEQ ID NO:115、116))在现有技术中有描述并且众所周知可催化一系列糖对糖反应,并且最近被描述为能够对大麻素糖苷进行糖对糖反应。然而,在现有技术中没有描述第三种酶(PL-152(Si94D_GA)(SEQ ID NO:145、146)),但在我们的筛选中发现其有效地进行糖对糖反应。在单个反应中组合多种糖基转移酶能够产生更多样化的大麻素糖苷,这些糖苷不是由单独表达的酶产生的。为了证明这一点,使用CBD和UDP-葡萄糖作为底物进行体外酶分析。将先前被证明可以产生CBD-1'-O-β-D-葡糖苷(OB1)的 PL-159(Pt88G_GA)与先前被证明可以将第二葡萄糖分子附连至 CBD-1'-O-β-D-葡糖苷(OB1)的葡萄糖部分的酶(PL-55(Sr76G1_GA)(SEQ ID NO:123、124),PL-32(OsEUGT11_GA)(SEQ ID NO:115、116),PL-152(Si94D_GA)(SEQ ID NO:145、146))组合。如前所述进行并分析体外测定。在现有技术中,Sr76G1被描述为能够将大麻素苷元转化为大麻素糖苷,而出人意料的是,我们未检测到使用大麻素苷元作为底物的这种酶的任何活性,但我们确实检测到了以大麻素糖苷为底物的活性。发现当与 Pt88G组合时,所有3种酶均可以将OB1转化为CBD-二-葡糖苷衍生物(OB2-4)。通过比较LC-MS/QTOF保留时间、测量质量和碎裂模式以及 cLogP,可以阐明Sr76G1、OsEUGT11和Si94D在具有不同键的糖反应中催化糖。Sr76G1被表明催化1→3葡萄糖-葡萄糖键(昆布二糖苷),而 OsEUGT11被表明催化1→4葡萄糖-葡萄糖键和1→6葡萄糖-葡萄糖键(龙胆二糖苷)。有意思的是,Si94D被表明以极高的效率(100%)催化1→6葡萄糖-葡萄糖键(龙胆二糖苷),如下表(表59)所示。结果最终表明Sr76G1对大麻素苷元没有活性,但实际上对葡萄糖分子有活性。催化具有不同键的糖- 糖反应的酶的发现极大地扩展了可以用糖基转移酶的不同组合产生的大麻素糖苷的多样性。
表61.通过将在大麻素苷元上有活性的糖基转移酶与在大麻素糖苷上有活性的糖基转移酶组合,将CBD体外酶促转化为具有不同糖键的多糖CBD- 葡糖苷。示出的是转化为每种各自产物的CBD量,以百分比表示。昆布二糖苷,具有1→3键的二-葡糖苷(OB2);龙胆二糖苷,具有1→6键的二-葡糖苷(OB3);纤维二糖苷,具有1→4键的二-萄糖苷(OB4)。ND;未检测到。
Figure BDA0003490979370002281
实施例21-酿酒酵母中大麻素和大麻素糖苷的毒性试验
众所周知,大麻素对微生物有毒性,并且人们认为这些化合物是由大麻植物产生的,作为抵御感染的防御机制。此外,越来越多的证据表明,各种大麻素是有效的抗微生物剂,已证明对一系列病原菌和真菌物种有效。产生大麻素的微生物菌株中的产物毒性将阻碍这些分子的高水平产生,糖基化这些分子可用于解毒它们并促进工程化微生物菌株的更高产生滴度。为了测量大麻素和大麻素糖苷的毒性作用,野生型酿酒酵母菌株BY4741 在补充有2%葡萄糖和溶于乙醇的不同浓度CBD和CBD-1'-O-β-D-葡糖基 3'-O-β-D-葡糖苷(OB6)的YP培养基中培养,调整浓度使所有细胞培养物中乙醇的终浓度为3%。将细胞接种到0.1的起始OD600并在30℃和200RPM 下孵育,并且72h后测量最终OD600。如下表60所示,增加溶液中CBD 的浓度导致最终OD600逐渐降低,而对于OB6,最终OD600在所有检测浓度下保持相对恒定。这表明虽然CBD对酵母具有毒性,但OB6在检测的浓度范围内是不具有毒性。
表62.在不同浓度的CBD和CBD-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷 (OB6)存在下培养的酿酒酵母的最终OD600。
浓度(μM)
Figure BDA0003490979370002291
序列表
<110> 奥克塔林生物制剂公司
<120> 产生糖基化大麻素的基因修饰的宿主细胞
<130> P19-002 WOPC
<150> EP19176773
<151> 2019-05-27
<160> 320
<170> PatentIn version 3.5
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gttactttga tcactactta cccaactgtt tctttggctg aaactcaaca cgtttctcac 180
ttcttgtctg cttacccaca agttactgaa aagagattcc acttgttgcc attcgaccca 240
aactctgcta acgctactga cccattcttc ttgagatggg aagctatcag aagatctgct 300
cacttgttgg ctccattgtt gtctccacca ttgtctgctt tgatcactga cgttactttg 360
atctctgctg ttttgccagt tactatcaac ttgcacttgc caaactacgt tttgttcact 420
gcttctgcta gaatgttctc tttgactgct tctttcccag ctatcgttgc ttctaagtct 480
acttcttctg gttctgttga attcgacgac gacttcatcg aaatcccagg tttgccacca 540
atcccattgt cttctgttcc accagctgtt atggactcta agtctttgtt cgctacttct 600
ttcttggaaa acggtaactc tttcgttaag tctaacggtg ttttgatcaa ctctttcgac 660
gctttggaag ctgacacttt ggttgctttg aacggtagaa gagttgttgc tggtttgcca 720
ccagtttacg ctgttggtcc attgttgcca tgtgaattcg aaaagagaga cgacccatct 780
acttctttga tcttgaagtg gttggacgac caaccagaag gttctgttgt ttacgtttct 840
ttcggttcta gattggcttt gtctatggaa caaactaagg aattgggtaa cggtttgttg 900
tcttctggtt gtagattctt gtgggttgtt aagggtaaga ctgttgacaa ggaagacgaa 960
gaatctttga agaacgtttt gggtcacgaa ttgatggaaa agatcaagga ccaaggtttg 1020
gttgttaaga actgggttga ccaagacaag gttttgtctc acagagctgt tggtggtttc 1080
gtttctcacg gtggttggaa ctctttggtt gaagctgcta gacacggtgt tccagttttg 1140
gtttggccac aattcggtga ccaaaagatc aacgctgaag ctgttgaatc tgctggtttg 1200
ggtatgtggg ttagatcttg gggttggggt actgaattga gagctaaggg tgacgaaatc 1260
ggtttgaaga tcaaggactt gatggctaac gacttcttga gagaacaagc taagagaatc 1320
gaagaagaag ctagaaaggc tatcggtgtt ggtggttctt ctgaaagaac tttcaaggaa 1380
ttgatcgaca agtggaagtg taacaacaac actcactag 1419
<210> 7
<211> 471
<212> PRT
<213> 水稻
<400> 7
Met Pro Ser Ser Gly Asp Ala Ala Gly Arg Arg Pro His Val Val Leu
1 5 10 15
Ile Pro Ser Ala Gly Met Gly His Leu Val Pro Phe Gly Arg Leu Ala
20 25 30
Val Ala Leu Ser Ser Gly His Gly Cys Asp Val Ser Leu Val Thr Val
35 40 45
Leu Pro Thr Val Ser Thr Ala Glu Ser Lys His Leu Asp Ala Leu Phe
50 55 60
Asp Ala Phe Pro Ala Val Arg Arg Leu Asp Phe Glu Leu Ala Pro Phe
65 70 75 80
Asp Ala Ser Glu Phe Pro Gly Ala Asp Pro Phe Phe Leu Arg Phe Glu
85 90 95
Ala Met Arg Arg Ser Ala Pro Leu Leu Gly Pro Leu Leu Thr Gly Ala
100 105 110
Gly Ala Ser Ala Leu Ala Thr Asp Ile Ala Leu Thr Ser Val Val Ile
115 120 125
Pro Val Ala Lys Glu Gln Gly Leu Pro Cys His Ile Leu Phe Thr Ala
130 135 140
Ser Ala Ala Met Leu Ser Leu Cys Ala Tyr Phe Pro Thr Tyr Leu Asp
145 150 155 160
Ala Asn Ala Gly Gly Gly Gly Gly Val Gly Asp Val Asp Ile Pro Gly
165 170 175
Val Tyr Arg Ile Pro Lys Ala Ser Ile Pro Gln Ala Leu His Asp Pro
180 185 190
Asn His Leu Phe Thr Arg Gln Phe Val Ala Asn Gly Arg Ser Leu Thr
195 200 205
Ser Ala Ala Gly Ile Leu Val Asn Thr Phe Asp Ala Leu Glu Pro Glu
210 215 220
Ala Val Ala Ala Leu Gln Gln Gly Lys Val Ala Ser Gly Phe Pro Pro
225 230 235 240
Val Phe Ala Val Gly Pro Leu Leu Pro Ala Ser Asn Gln Ala Lys Asp
245 250 255
Pro Gln Ala Asn Tyr Met Glu Trp Leu Asp Ala Gln Pro Ala Arg Ser
260 265 270
Val Val Tyr Val Ser Phe Gly Ser Arg Lys Ala Ile Ser Arg Glu Gln
275 280 285
Leu Arg Glu Leu Ala Ala Gly Leu Glu Gly Ser Gly His Arg Phe Leu
290 295 300
Trp Val Val Lys Ser Thr Val Val Asp Arg Asp Asp Ala Ala Glu Leu
305 310 315 320
Gly Glu Leu Leu Asp Glu Gly Phe Leu Glu Arg Val Glu Lys Arg Gly
325 330 335
Leu Val Thr Lys Ala Trp Val Asp Gln Glu Glu Val Leu Lys His Glu
340 345 350
Ser Val Ala Leu Phe Val Ser His Cys Gly Trp Asn Ser Val Thr Glu
355 360 365
Ala Ala Ala Ser Gly Val Pro Val Leu Ala Leu Pro Arg Phe Gly Asp
370 375 380
Gln Arg Val Asn Ser Gly Val Val Ala Arg Ala Gly Leu Gly Val Trp
385 390 395 400
Ala Asp Thr Trp Ser Trp Glu Gly Glu Ala Gly Val Ile Gly Ala Glu
405 410 415
Glu Ile Ser Glu Lys Val Lys Ala Ala Met Ala Asp Glu Ala Leu Arg
420 425 430
Met Lys Ala Ala Ser Leu Ala Glu Ala Ala Ala Lys Ala Val Ala Gly
435 440 445
Gly Gly Ser Ser His Arg Cys Leu Ala Glu Phe Ala Arg Leu Cys Gln
450 455 460
Gly Gly Thr Cys Arg Thr Asn
465 470
<210> 8
<211> 1416
<212> DNA
<213> 水稻
<400> 8
atgccatctt ctggtgacgc tgctggtaga agaccacacg ttgttttgat cccatctgct 60
ggtatgggtc acttggttcc attcggtaga ttggctgttg ctttgtcttc tggtcacggt 120
tgtgacgttt ctttggttac tgttttgcca actgtttcta ctgctgaatc taagcacttg 180
gacgctttgt tcgacgcttt cccagctgtt agaagattgg acttcgaatt ggctccattc 240
gacgcttctg aattcccagg tgctgaccca ttcttcttga gattcgaagc tatgagaaga 300
tctgctccat tgttgggtcc attgttgact ggtgctggtg cttctgcttt ggctactgac 360
atcgctttga cttctgttgt tatcccagtt gctaaggaac aaggtttgcc atgtcacatc 420
ttgttcactg cttctgctgc tatgttgtct ttgtgtgctt acttcccaac ttacttggac 480
gctaacgctg gtggtggtgg tggtgttggt gacgttgaca tcccaggtgt ttacagaatc 540
ccaaaggctt ctatcccaca agctttgcac gacccaaacc acttgttcac tagacaattc 600
gttgctaacg gtagatcttt gacttctgct gctggtatct tggttaacac tttcgacgct 660
ttggaaccag aagctgttgc tgctttgcaa caaggtaagg ttgcttctgg tttcccacca 720
gttttcgctg ttggtccatt gttgccagct tctaaccaag ctaaggaccc acaagctaac 780
tacatggaat ggttggacgc tcaaccagct agatctgttg tttacgtttc tttcggttct 840
agaaaggcta tctctagaga acaattgaga gaattggctg ctggtttgga aggttctggt 900
cacagattct tgtgggttgt taagtctact gttgttgaca gagacgacgc tgctgaattg 960
ggtgaattgt tggacgaagg tttcttggaa agagttgaaa agagaggttt ggttactaag 1020
gcttgggttg accaagaaga agttttgaag cacgaatctg ttgctttgtt cgtttctcac 1080
tgtggttgga actctgttac tgaagctgct gcttctggtg ttccagtttt ggctttgcca 1140
agattcggtg accaaagagt taactctggt gttgttgcta gagctggttt gggtgtttgg 1200
gctgacactt ggtcttggga aggtgaagct ggtgttatcg gtgctgaaga aatctctgaa 1260
aaggttaagg ctgctatggc tgacgaagct ttgagaatga aggctgcttc tttggctgaa 1320
gctgctgcta aggctgttgc tggtggtggt tcttctcaca gatgtttggc tgaattcgct 1380
agattgtgtc aaggtggtac ttgtagaact aactag 1416
<210> 9
<211> 457
<212> PRT
<213> 甜荞(Fagopyrum esculentum)
<400> 9
Met Met Gly Asp Leu Thr Thr Ser Phe Pro Ala Thr Thr Leu Thr Thr
1 5 10 15
Asn Asp Gln Pro His Val Val Val Cys Ser Gly Ala Gly Met Gly His
20 25 30
Leu Thr Pro Phe Leu Asn Leu Ala Ser Ala Leu Ser Ser Ala Pro Tyr
35 40 45
Asn Cys Lys Val Thr Leu Leu Ile Val Ile Pro Leu Ile Thr Asp Ala
50 55 60
Glu Ser His His Ile Ser Ser Phe Phe Ser Ser His Pro Thr Ile His
65 70 75 80
Arg Leu Asp Phe His Val Asn Leu Pro Ala Pro Lys Pro Asn Val Asp
85 90 95
Pro Phe Phe Leu Arg Tyr Lys Ser Ile Ser Asp Ser Ala His Arg Leu
100 105 110
Pro Val His Leu Ser Ala Leu Ser Pro Pro Ile Ser Ala Val Phe Ser
115 120 125
Asp Phe Leu Phe Thr Gln Gly Leu Asn Thr Thr Leu Pro His Leu Pro
130 135 140
Asn Tyr Thr Phe Thr Thr Thr Ser Ala Arg Phe Phe Thr Leu Met Ser
145 150 155 160
Tyr Val Pro His Leu Ala Lys Ser Ser Ser Ser Ser Pro Val Glu Ile
165 170 175
Pro Gly Leu Glu Pro Phe Pro Thr Asp Asn Ile Pro Pro Pro Phe Phe
180 185 190
Asn Pro Glu His Ile Phe Thr Ser Phe Thr Ile Ser Asn Ala Lys Tyr
195 200 205
Phe Ser Leu Ser Lys Gly Ile Leu Val Asn Thr Phe Asp Ser Phe Glu
210 215 220
Pro Glu Thr Leu Ser Ala Leu Asn Ser Gly Asp Thr Leu Ser Asp Leu
225 230 235 240
Pro Pro Val Ile Pro Ile Gly Pro Leu Asn Glu Leu Glu His Asn Lys
245 250 255
Gln Glu Glu Leu Leu Pro Trp Leu Asp Gln Gln Pro Glu Lys Ser Val
260 265 270
Leu Tyr Val Ser Phe Gly Asn Arg Thr Ala Met Ser Ser Asp Gln Ile
275 280 285
Leu Glu Leu Gly Met Gly Leu Glu Arg Ser Asp Cys Arg Phe Ile Trp
290 295 300
Val Val Lys Thr Ser Lys Ile Asp Lys Asp Asp Lys Ser Glu Leu Arg
305 310 315 320
Lys Leu Phe Gly Glu Glu Leu Tyr Leu Lys Leu Ser Glu Lys Gly Lys
325 330 335
Leu Val Lys Trp Val Asn Gln Thr Glu Ile Leu Gly His Thr Ala Val
340 345 350
Gly Gly Phe Leu Ser His Cys Gly Trp Asn Ser Val Met Glu Ala Ala
355 360 365
Arg Arg Gly Val Pro Ile Leu Ala Trp Pro Gln His Gly Asp Gln Arg
370 375 380
Glu Asn Ala Trp Val Val Glu Lys Ala Gly Leu Gly Val Trp Glu Arg
385 390 395 400
Glu Trp Ala Ser Gly Ile Gln Ala Ala Ile Val Glu Lys Val Lys Met
405 410 415
Ile Met Gly Asn Asn Asp Leu Arg Lys Ser Ala Met Lys Val Gly Glu
420 425 430
Glu Ala Lys Arg Ala Cys Asp Val Gly Gly Ser Ser Ala Thr Ala Leu
435 440 445
Met Asn Ile Ile Gly Ser Leu Lys Arg
450 455
<210> 10
<211> 1374
<212> DNA
<213> 甜荞
<400> 10
atgatgggtg acttgactac ttctttccca gctactactt tgactactaa cgaccaacca 60
cacgttgttg tttgttctgg tgctggtatg ggtcacttga ctccattctt gaacttggct 120
tctgctttgt cttctgctcc atacaactgt aaggttactt tgttgatcgt tatcccattg 180
atcactgacg ctgaatctca ccacatctct tctttcttct cttctcaccc aactatccac 240
agattggact tccacgttaa cttgccagct ccaaagccaa acgttgaccc attcttcttg 300
agatacaagt ctatctctga ctctgctcac agattgccag ttcacttgtc tgctttgtct 360
ccaccaatct ctgctgtttt ctctgacttc ttgttcactc aaggtttgaa cactactttg 420
ccacacttgc caaactacac tttcactact acttctgcta gattcttcac tttgatgtct 480
tacgttccac acttggctaa gtcttcttct tcttctccag ttgaaatccc aggtttggaa 540
ccattcccaa ctgacaacat cccaccacca ttcttcaacc cagaacacat cttcacttct 600
ttcactatct ctaacgctaa gtacttctct ttgtctaagg gtatcttggt taacactttc 660
gactctttcg aaccagaaac tttgtctgct ttgaactctg gtgacacttt gtctgacttg 720
ccaccagtta tcccaatcgg tccattgaac gaattggaac acaacaagca agaagaattg 780
ttgccatggt tggaccaaca accagaaaag tctgttttgt acgtttcttt cggtaacaga 840
actgctatgt cttctgacca aatcttggaa ttgggtatgg gtttggaaag atctgactgt 900
agattcatct gggttgttaa gacttctaag atcgacaagg acgacaagtc tgaattgaga 960
aagttgttcg gtgaagaatt gtacttgaag ttgtctgaaa agggtaagtt ggttaagtgg 1020
gttaaccaaa ctgaaatctt gggtcacact gctgttggtg gtttcttgtc tcactgtggt 1080
tggaactctg ttatggaagc tgctagaaga ggtgttccaa tcttggcttg gccacaacac 1140
ggtgaccaaa gagaaaacgc ttgggttgtt gaaaaggctg gtttgggtgt ttgggaaaga 1200
gaatgggctt ctggtatcca agctgctatc gttgaaaagg ttaagatgat catgggtaac 1260
aacgacttga gaaagtctgc tatgaaggtt ggtgaagaag ctaagagagc ttgtgacgtt 1320
ggtggttctt ctgctactgc tttgatgaac atcatcggtt ctttgaagag atag 1374
<210> 11
<211> 480
<212> PRT
<213> 大豆(Glycine max)
<400> 11
Met Ser Ser Ser Glu Gly Val Val His Val Ala Phe Leu Pro Ser Ala
1 5 10 15
Gly Met Gly His Leu Asn Pro Phe Leu Arg Leu Ala Ala Thr Phe Ile
20 25 30
Arg Tyr Gly Cys Lys Val Thr Leu Ile Thr Pro Lys Pro Thr Val Ser
35 40 45
Leu Ala Glu Ser Asn Leu Ile Ser Arg Phe Cys Ser Ser Phe Pro His
50 55 60
Gln Val Thr Gln Leu Asp Leu Asn Leu Val Ser Val Asp Pro Thr Thr
65 70 75 80
Val Asp Thr Ile Asp Pro Phe Phe Leu Gln Phe Glu Thr Ile Arg Arg
85 90 95
Ser Leu His Leu Leu Pro Pro Ile Leu Ser Leu Leu Ser Thr Pro Leu
100 105 110
Ser Ala Phe Ile Tyr Asp Ile Thr Leu Ile Thr Pro Leu Leu Ser Val
115 120 125
Ile Glu Lys Leu Ser Cys Pro Ser Tyr Leu Tyr Phe Thr Ser Ser Ala
130 135 140
Arg Met Phe Ser Phe Phe Ala Arg Val Ser Val Leu Ser Ala Ser Asn
145 150 155 160
Pro Gly Gln Thr Pro Ser Ser Phe Ile Gly Asp Asp Gly Val Lys Ile
165 170 175
Pro Gly Phe Thr Ser Pro Ile Pro Arg Ser Ser Val Pro Pro Ala Ile
180 185 190
Leu Gln Ala Ser Ser Asn Leu Phe Gln Arg Ile Met Leu Glu Asp Ser
195 200 205
Ala Asn Val Thr Lys Leu Asn Asn Gly Val Phe Ile Asn Ser Phe Glu
210 215 220
Glu Leu Glu Gly Glu Ala Leu Ala Ala Leu Asn Gly Gly Lys Val Leu
225 230 235 240
Glu Gly Leu Pro Pro Val Tyr Gly Val Gly Pro Leu Met Ala Cys Glu
245 250 255
Tyr Glu Lys Gly Asp Glu Glu Gly Gln Lys Gly Cys Met Ser Ser Ile
260 265 270
Val Lys Trp Leu Asp Glu Gln Ser Lys Gly Ser Val Val Tyr Val Ser
275 280 285
Leu Gly Asn Arg Thr Glu Thr Arg Arg Glu Gln Ile Lys Asp Met Ala
290 295 300
Leu Gly Leu Ile Glu Cys Gly Tyr Gly Phe Leu Trp Val Val Lys Leu
305 310 315 320
Lys Arg Val Asp Lys Glu Asp Glu Glu Gly Leu Glu Glu Val Leu Gly
325 330 335
Ser Glu Leu Ser Ser Lys Val Lys Glu Lys Gly Val Val Val Lys Glu
340 345 350
Phe Val Asp Gln Val Glu Ile Leu Gly His Pro Ser Val Gly Gly Phe
355 360 365
Leu Ser His Gly Gly Trp Asn Ser Val Thr Glu Thr Val Trp Lys Gly
370 375 380
Val Pro Cys Leu Ser Trp Pro Gln His Ser Asp Gln Lys Met Ser Ala
385 390 395 400
Glu Val Ile Arg Met Ser Gly Met Gly Ile Trp Pro Glu Glu Trp Gly
405 410 415
Trp Gly Thr Gln Asp Val Val Lys Gly Asp Glu Ile Ala Lys Arg Ile
420 425 430
Lys Glu Met Met Ser Asn Glu Ser Leu Arg Val Lys Ala Gly Glu Leu
435 440 445
Lys Glu Ala Ala Leu Lys Ala Ala Gly Val Gly Gly Ser Cys Glu Val
450 455 460
Thr Ile Lys Arg Gln Ile Glu Glu Trp Lys Arg Asn Ala Gln Ala Asn
465 470 475 480
<210> 12
<211> 1443
<212> DNA
<213> 大豆
<400> 12
atgtcttctt ctgaaggtgt tgttcacgtt gctttcttgc catctgctgg tatgggtcac 60
ttgaacccat tcttgagatt ggctgctact ttcatcagat acggttgtaa ggttactttg 120
atcactccaa agccaactgt ttctttggct gaatctaact tgatctctag attctgttct 180
tctttcccac accaagttac tcaattggac ttgaacttgg tttctgttga cccaactact 240
gttgacacta tcgacccatt cttcttgcaa ttcgaaacta tcagaagatc tttgcacttg 300
ttgccaccaa tcttgtcttt gttgtctact ccattgtctg ctttcatcta cgacatcact 360
ttgatcactc cattgttgtc tgttatcgaa aagttgtctt gtccatctta cttgtacttc 420
acttcttctg ctagaatgtt ctctttcttc gctagagttt ctgttttgtc tgcttctaac 480
ccaggtcaaa ctccatcttc tttcatcggt gacgacggtg ttaagatccc aggtttcact 540
tctccaatcc caagatcttc tgttccacca gctatcttgc aagcttcttc taacttgttc 600
caaagaatca tgttggaaga ctctgctaac gttactaagt tgaacaacgg tgttttcatc 660
aactctttcg aagaattgga aggtgaagct ttggctgctt tgaacggtgg taaggttttg 720
gaaggtttgc caccagttta cggtgttggt ccattgatgg cttgtgaata cgaaaagggt 780
gacgaagaag gtcaaaaggg ttgtatgtct tctatcgtta agtggttgga cgaacaatct 840
aagggttctg ttgtttacgt ttctttgggt aacagaactg aaactagaag agaacaaatc 900
aaggacatgg ctttgggttt gatcgaatgt ggttacggtt tcttgtgggt tgttaagttg 960
aagagagttg acaaggaaga cgaagaaggt ttggaagaag ttttgggttc tgaattgtct 1020
tctaaggtta aggaaaaggg tgttgttgtt aaggaattcg ttgaccaagt tgaaatcttg 1080
ggtcacccat ctgttggtgg tttcttgtct cacggtggtt ggaactctgt tactgaaact 1140
gtttggaagg gtgttccatg tttgtcttgg ccacaacact ctgaccaaaa gatgtctgct 1200
gaagttatca gaatgtctgg tatgggtatc tggccagaag aatggggttg gggtactcaa 1260
gacgttgtta agggtgacga aatcgctaag agaatcaagg aaatgatgtc taacgaatct 1320
ttgagagtta aggctggtga attgaaggaa gctgctttga aggctgctgg tgttggtggt 1380
tcttgtgaag ttactatcaa gagacaaatc gaagaatgga agagaaacgc tcaagctaac 1440
tag 1443
<210> 13
<211> 475
<212> PRT
<213> 玉蜀黍
<400> 13
Met Ala Ala Asn Gly Gly Asp His Thr Ser Ala Arg Pro His Val Val
1 5 10 15
Leu Leu Pro Ser Ala Gly Met Gly His Leu Val Pro Phe Ala Arg Leu
20 25 30
Ala Val Ala Leu Ser Glu Gly His Gly Cys Asn Val Ser Val Ala Ala
35 40 45
Val Gln Pro Thr Val Ser Ser Ala Glu Ser Arg Leu Leu Asp Ala Leu
50 55 60
Phe Val Ala Ala Ala Pro Ala Val Arg Arg Leu Asp Phe Arg Leu Ala
65 70 75 80
Pro Phe Asp Glu Ser Glu Phe Pro Gly Ala Asp Pro Phe Phe Leu Arg
85 90 95
Phe Glu Ala Thr Arg Arg Ser Ala Pro Leu Leu Gly Pro Leu Leu Asp
100 105 110
Ala Ala Glu Ala Ser Ala Leu Val Thr Asp Ile Val Leu Ala Ser Val
115 120 125
Ala Leu Pro Val Ala Arg Glu Arg Gly Val Pro Cys Tyr Val Leu Phe
130 135 140
Thr Ser Ser Ala Ala Met Leu Ser Leu Cys Ala Tyr Phe Pro Ala Tyr
145 150 155 160
Leu Asp Ala His Ala Ala Ala Gly Ser Val Gly Val Gly Val Gly Asn
165 170 175
Val Asp Ile Pro Gly Val Phe Arg Ile Pro Lys Ser Ser Val Pro Gln
180 185 190
Ala Leu His Asp Pro Asp His Leu Phe Thr Gln Gln Phe Val Ala Asn
195 200 205
Gly Arg Cys Leu Val Ala Cys Asp Gly Ile Leu Val Asn Thr Phe Asp
210 215 220
Ala Phe Glu Pro Asp Ala Val Thr Ala Leu Arg Gln Gly Ser Ile Thr
225 230 235 240
Val Ser Gly Gly Phe Pro Pro Val Phe Thr Val Gly Pro Met Leu Pro
245 250 255
Val Arg Phe Gln Ala Glu Glu Thr Ala Asp Tyr Met Arg Trp Leu Ser
260 265 270
Ala Gln Pro Pro Arg Ser Val Val Tyr Val Ser Phe Gly Ser Arg Lys
275 280 285
Ala Ile Pro Arg Asp Gln Leu Arg Glu Leu Ala Ala Gly Leu Glu Ala
290 295 300
Ser Gly Lys Arg Phe Leu Trp Val Val Lys Ser Thr Ile Val Asp Arg
305 310 315 320
Asp Asp Thr Ala Asp Leu Gly Gly Leu Leu Gly Asp Gly Phe Leu Glu
325 330 335
Arg Val Gln Gly Arg Ala Phe Val Thr Met Gly Trp Val Glu Gln Glu
340 345 350
Glu Ile Leu Gln His Gly Ser Val Gly Leu Phe Ile Ser His Cys Gly
355 360 365
Trp Asn Ser Leu Thr Glu Ala Ala Ala Phe Gly Val Pro Val Leu Ala
370 375 380
Trp Pro Arg Phe Gly Asp Gln Arg Val Asn Ala Ala Leu Val Ala Arg
385 390 395 400
Ser Gly Leu Gly Ala Trp Glu Glu Gly Trp Thr Trp Asp Gly Glu Glu
405 410 415
Gly Leu Thr Thr Arg Lys Glu Val Ala Lys Lys Ile Lys Gly Met Met
420 425 430
Gly Tyr Asp Ala Val Ala Glu Lys Ala Ala Lys Val Gly Asp Ala Ala
435 440 445
Ala Ala Ala Ile Ala Lys Cys Gly Thr Ser Tyr Gln Ser Leu Glu Glu
450 455 460
Phe Val Gln Arg Cys Arg Asp Ala Glu Arg Lys
465 470 475
<210> 14
<211> 1428
<212> DNA
<213> 玉蜀黍
<400> 14
atggctgcta acggtggtga ccacacttct gctagaccac acgttgtttt gttgccatct 60
gctggtatgg gtcacttggt tccattcgct agattggctg ttgctttgtc tgaaggtcac 120
ggttgtaacg tttctgttgc tgctgttcaa ccaactgttt cttctgctga atctagattg 180
ttggacgctt tgttcgttgc tgctgctcca gctgttagaa gattggactt cagattggct 240
ccattcgacg aatctgaatt cccaggtgct gacccattct tcttgagatt cgaagctact 300
agaagatctg ctccattgtt gggtccattg ttggacgctg ctgaagcttc tgctttggtt 360
actgacatcg ttttggcttc tgttgctttg ccagttgcta gagaaagagg tgttccatgt 420
tacgttttgt tcacttcttc tgctgctatg ttgtctttgt gtgcttactt cccagcttac 480
ttggacgctc acgctgctgc tggttctgtt ggtgttggtg ttggtaacgt tgacatccca 540
ggtgttttca gaatcccaaa gtcttctgtt ccacaagctt tgcacgaccc agaccacttg 600
ttcactcaac aattcgttgc taacggtaga tgtttggttg cttgtgacgg tatcttggtt 660
aacactttcg acgctttcga accagacgct gttactgctt tgagacaagg ttctatcact 720
gtttctggtg gtttcccacc agttttcact gttggtccaa tgttgccagt tagattccaa 780
gctgaagaaa ctgctgacta catgagatgg ttgtctgctc aaccaccaag atctgttgtt 840
tacgtttctt tcggttctag aaaggctatc ccaagagacc aattgagaga attggctgct 900
ggtttggaag cttctggtaa gagattcttg tgggttgtta agtctactat cgttgacaga 960
gacgacactg ctgacttggg tggtttgttg ggtgacggtt tcttggaaag agttcaaggt 1020
agagctttcg ttactatggg ttgggttgaa caagaagaaa tcttgcaaca cggttctgtt 1080
ggtttgttca tctctcactg tggttggaac tctttgactg aagctgctgc tttcggtgtt 1140
ccagttttgg cttggccaag attcggtgac caaagagtta acgctgcttt ggttgctaga 1200
tctggtttgg gtgcttggga agaaggttgg acttgggacg gtgaagaagg tttgactact 1260
agaaaggaag ttgctaagaa gatcaagggt atgatgggtt acgacgctgt tgctgaaaag 1320
gctgctaagg ttggtgacgc tgctgctgct gctatcgcta agtgtggtac ttcttaccaa 1380
tctttggaag aattcgttca aagatgtaga gacgctgaaa gaaagtag 1428
<210> 15
<211> 470
<212> PRT
<213> 芒果(Mangifera indica)
<400> 15
Met Ser Ala Ser Asp Ala Leu Asn Ser Cys Pro His Val Ala Leu Leu
1 5 10 15
Leu Ser Ser Gly Met Gly His Leu Thr Pro Cys Leu Arg Phe Ala Ala
20 25 30
Thr Leu Val Gln His His Cys Arg Val Thr Ile Ile Thr Asn Tyr Pro
35 40 45
Thr Val Ser Val Ala Glu Ser Arg Ala Ile Ser Leu Leu Leu Ser Asp
50 55 60
Phe Pro Gln Ile Thr Glu Lys Gln Phe His Leu Leu Pro Phe Asp Pro
65 70 75 80
Ser Thr Ala Asn Thr Thr Asp Pro Phe Phe Leu Arg Trp Glu Ala Ile
85 90 95
Arg Arg Ser Ala His Leu Leu Asn Pro Leu Leu Ser Ser Ile Ser Pro
100 105 110
Pro Leu Ser Ala Leu Val Ile Asp Ser Ser Leu Val Ser Ser Phe Val
115 120 125
Pro Val Ala Ala Asn Leu Asp Leu Pro Ser Tyr Val Leu Phe Thr Ser
130 135 140
Ser Thr Arg Met Cys Ser Leu Glu Glu Thr Phe Pro Ala Phe Val Ala
145 150 155 160
Ser Lys Thr Asn Phe Asp Ser Ile Gln Leu Asp Asp Val Ile Glu Ile
165 170 175
Pro Gly Phe Ser Pro Val Pro Val Ser Ser Val Pro Pro Val Phe Leu
180 185 190
Asn Leu Asn His Leu Phe Thr Thr Met Leu Ile Gln Asn Gly Gln Ser
195 200 205
Phe Arg Lys Ala Asn Gly Ile Leu Ile Asn Thr Phe Glu Ala Leu Glu
210 215 220
Gly Gly Ile Leu Pro Gly Ile Asn Asp Lys Arg Ala Ala Asp Gly Leu
225 230 235 240
Pro Pro Tyr Cys Ser Val Gly Pro Leu Leu Pro Cys Lys Phe Glu Lys
245 250 255
Thr Glu Cys Ser Ala Pro Val Lys Trp Leu Asp Asp Gln Pro Glu Gly
260 265 270
Ser Val Val Tyr Val Ser Phe Gly Ser Arg Phe Ala Leu Ser Ser Glu
275 280 285
Gln Ile Lys Glu Leu Gly Asp Gly Leu Ile Arg Ser Gly Cys Arg Phe
290 295 300
Leu Trp Val Val Lys Cys Lys Lys Val Asp Gln Glu Asp Glu Glu Ser
305 310 315 320
Leu Asp Glu Leu Leu Gly Arg Asp Val Leu Glu Lys Ile Lys Lys Tyr
325 330 335
Gly Phe Val Ile Lys Asn Trp Val Asn Gln Gln Glu Ile Leu Asp His
340 345 350
Arg Ala Val Gly Gly Phe Val Thr His Gly Gly Trp Asn Ser Ser Met
355 360 365
Glu Ala Val Trp His Gly Val Pro Met Leu Val Trp Pro Gln Phe Gly
370 375 380
Asp Gln Lys Ile Asn Ala Glu Val Ile Glu Arg Ser Gly Leu Gly Met
385 390 395 400
Trp Val Lys Arg Trp Gly Trp Gly Thr Gln Gln Leu Val Lys Gly Glu
405 410 415
Glu Ile Gly Glu Arg Ile Lys Asp Leu Met Gly Asn Asn Pro Leu Arg
420 425 430
Val Arg Ala Lys Thr Leu Arg Glu Glu Ala Arg Lys Ala Ile Glu Val
435 440 445
Gly Gly Ser Ser Glu Lys Thr Leu Lys Glu Leu Ile Glu Asn Trp Lys
450 455 460
Lys Thr Ser Arg Lys Thr
465 470
<210> 16
<211> 1413
<212> DNA
<213> 芒果
<400> 16
atgtctgctt ctgacgcttt gaactcttgt ccacacgttg ctttgttgtt gtcttctggt 60
atgggtcact tgactccatg tttgagattc gctgctactt tggttcaaca ccactgtaga 120
gttactatca tcactaacta cccaactgtt tctgttgctg aatctagagc tatctctttg 180
ttgttgtctg acttcccaca aatcactgaa aagcaattcc acttgttgcc attcgaccca 240
tctactgcta acactactga cccattcttc ttgagatggg aagctatcag aagatctgct 300
cacttgttga acccattgtt gtcttctatc tctccaccat tgtctgcttt ggttatcgac 360
tcttctttgg tttcttcttt cgttccagtt gctgctaact tggacttgcc atcttacgtt 420
ttgttcactt cttctactag aatgtgttct ttggaagaaa ctttcccagc tttcgttgct 480
tctaagacta acttcgactc tatccaattg gacgacgtta tcgaaatccc aggtttctct 540
ccagttccag tttcttctgt tccaccagtt ttcttgaact tgaaccactt gttcactact 600
atgttgatcc aaaacggtca atctttcaga aaggctaacg gtatcttgat caacactttc 660
gaagctttgg aaggtggtat cttgccaggt atcaacgaca agagagctgc tgacggtttg 720
ccaccatact gttctgttgg tccattgttg ccatgtaagt tcgaaaagac tgaatgttct 780
gctccagtta agtggttgga cgaccaacca gaaggttctg ttgtttacgt ttctttcggt 840
tctagattcg ctttgtcttc tgaacaaatc aaggaattgg gtgacggttt gatcagatct 900
ggttgtagat tcttgtgggt tgttaagtgt aagaaggttg accaagaaga cgaagaatct 960
ttggacgaat tgttgggtag agacgttttg gaaaagatca agaagtacgg tttcgttatc 1020
aagaactggg ttaaccaaca agaaatcttg gaccacagag ctgttggtgg tttcgttact 1080
cacggtggtt ggaactcttc tatggaagct gtttggcacg gtgttccaat gttggtttgg 1140
ccacaattcg gtgaccaaaa gatcaacgct gaagttatcg aaagatctgg tttgggtatg 1200
tgggttaaga gatggggttg gggtactcaa caattggtta agggtgaaga aatcggtgaa 1260
agaatcaagg acttgatggg taacaaccca ttgagagtta gagctaagac tttgagagaa 1320
gaagctagaa aggctatcga agttggtggt tcttctgaaa agactttgaa ggaattgatc 1380
gaaaactgga agaagacttc tagaaagact tag 1413
<210> 17
<211> 477
<212> PRT
<213> 三花龙胆(Gentiana triflora)
<400> 17
Met Gly Ser Leu Thr Asn Asn Asp Asn Leu His Ile Phe Leu Val Cys
1 5 10 15
Phe Ile Gly Gln Gly Val Val Asn Pro Met Leu Arg Leu Gly Lys Ala
20 25 30
Phe Ala Ser Lys Gly Leu Leu Val Thr Leu Ser Ala Pro Glu Ile Val
35 40 45
Gly Thr Glu Ile Arg Lys Ala Asn Asn Leu Asn Asp Asp Gln Pro Ile
50 55 60
Lys Val Gly Ser Gly Met Ile Arg Phe Glu Phe Phe Asp Asp Gly Trp
65 70 75 80
Glu Ser Val Asn Gly Ser Lys Pro Phe Asp Val Trp Val Tyr Ile Asn
85 90 95
His Leu Asp Gln Thr Gly Arg Gln Lys Leu Pro Ile Met Leu Lys Lys
100 105 110
His Glu Glu Thr Gly Thr Pro Val Ser Cys Leu Ile Leu Asn Pro Leu
115 120 125
Val Pro Trp Val Ala Asp Val Ala Asp Ser Leu Gln Ile Pro Cys Ala
130 135 140
Thr Leu Trp Val Gln Ser Cys Ala Ser Phe Ser Ala Tyr Tyr His Tyr
145 150 155 160
His His Gly Leu Val Pro Phe Pro Thr Glu Ser Glu Pro Glu Ile Asp
165 170 175
Val Gln Leu Pro Gly Met Pro Leu Leu Lys Tyr Asp Glu Val Pro Asp
180 185 190
Tyr Leu His Pro Arg Thr Pro Tyr Pro Phe Phe Gly Thr Asn Ile Leu
195 200 205
Gly Gln Phe Lys Asn Leu Ser Lys Asn Phe Cys Ile Leu Met Asp Thr
210 215 220
Phe Tyr Glu Leu Glu His Glu Ile Ile Asp Asn Met Cys Lys Leu Cys
225 230 235 240
Pro Ile Lys Pro Ile Gly Pro Leu Phe Lys Ile Pro Lys Asp Pro Ser
245 250 255
Ser Asn Gly Ile Thr Gly Asn Phe Met Lys Val Asp Asp Cys Lys Glu
260 265 270
Trp Leu Asp Ser Arg Pro Thr Ser Thr Val Val Tyr Val Ser Val Gly
275 280 285
Ser Val Val Tyr Leu Lys Gln Glu Gln Val Thr Glu Met Ala Tyr Gly
290 295 300
Ile Leu Asn Ser Glu Val Ser Phe Leu Trp Val Leu Arg Pro Pro Ser
305 310 315 320
Lys Arg Ile Gly Thr Glu Pro His Val Leu Pro Glu Glu Phe Trp Glu
325 330 335
Lys Ala Gly Asp Arg Gly Lys Val Val Gln Trp Ser Pro Gln Glu Gln
340 345 350
Val Leu Ala His Pro Ala Thr Val Gly Phe Leu Thr His Cys Gly Trp
355 360 365
Asn Ser Thr Gln Glu Ala Ile Ser Ser Gly Val Pro Val Ile Thr Phe
370 375 380
Pro Gln Phe Gly Asp Gln Val Thr Asn Ala Lys Phe Leu Val Glu Glu
385 390 395 400
Phe Lys Val Gly Val Arg Leu Gly Arg Gly Glu Leu Glu Asn Arg Ile
405 410 415
Ile Thr Arg Asp Glu Val Glu Arg Ala Leu Arg Glu Ile Thr Ser Gly
420 425 430
Pro Lys Ala Glu Glu Val Lys Glu Asn Ala Leu Lys Trp Lys Lys Lys
435 440 445
Ala Glu Glu Thr Val Ala Lys Gly Gly Tyr Ser Glu Arg Asn Leu Val
450 455 460
Gly Phe Ile Glu Glu Val Ala Arg Lys Thr Gly Thr Lys
465 470 475
<210> 18
<211> 1434
<212> DNA
<213> 三花龙胆
<400> 18
atgggttctt tgactaacaa cgacaacttg cacatcttct tggtttgttt catcggtcaa 60
ggtgttgtta acccaatgtt gagattgggt aaggctttcg cttctaaggg tttgttggtt 120
actttgtctg ctccagaaat cgttggtact gaaatcagaa aggctaacaa cttgaacgac 180
gaccaaccaa tcaaggttgg ttctggtatg atcagattcg aattcttcga cgacggttgg 240
gaatctgtta acggttctaa gccattcgac gtttgggttt acatcaacca cttggaccaa 300
actggtagac aaaagttgcc aatcatgttg aagaagcacg aagaaactgg tactccagtt 360
tcttgtttga tcttgaaccc attggttcca tgggttgctg acgttgctga ctctttgcaa 420
atcccatgtg ctactttgtg ggttcaatct tgtgcttctt tctctgctta ctaccactac 480
caccacggtt tggttccatt cccaactgaa tctgaaccag aaatcgacgt tcaattgcca 540
ggtatgccat tgttgaagta cgacgaagtt ccagactact tgcacccaag aactccatac 600
ccattcttcg gtactaacat cttgggtcaa ttcaagaact tgtctaagaa cttctgtatc 660
ttgatggaca ctttctacga attggaacac gaaatcatcg acaacatgtg taagttgtgt 720
ccaatcaagc caatcggtcc attgttcaag atcccaaagg acccatcttc taacggtatc 780
actggtaact tcatgaaggt tgacgactgt aaggaatggt tggactctag accaacttct 840
actgttgttt acgtttctgt tggttctgtt gtttacttga agcaagaaca agttactgaa 900
atggcttacg gtatcttgaa ctctgaagtt tctttcttgt gggttttgag accaccatct 960
aagagaatcg gtactgaacc acacgttttg ccagaagaat tctgggaaaa ggctggtgac 1020
agaggtaagg ttgttcaatg gtctccacaa gaacaagttt tggctcaccc agctactgtt 1080
ggtttcttga ctcactgtgg ttggaactct actcaagaag ctatctcttc tggtgttcca 1140
gttatcactt tcccacaatt cggtgaccaa gttactaacg ctaagttctt ggttgaagaa 1200
ttcaaggttg gtgttagatt gggtagaggt gaattggaaa acagaatcat cactagagac 1260
gaagttgaaa gagctttgag agaaatcact tctggtccaa aggctgaaga agttaaggaa 1320
aacgctttga agtggaagaa gaaggctgaa gaaactgttg ctaagggtgg ttactctgaa 1380
agaaacttgg ttggtttcat cgaagaagtt gctagaaaga ctggtactaa gtag 1434
<210> 19
<211> 515
<212> PRT
<213> 胭脂虫
<400> 19
Met Glu Phe Arg Leu Leu Ile Leu Ala Leu Phe Ser Val Leu Met Ser
1 5 10 15
Thr Ser Asn Gly Ala Glu Ile Leu Ala Leu Phe Pro Ile His Gly Ile
20 25 30
Ser Asn Tyr Asn Val Ala Glu Ala Leu Leu Lys Thr Leu Ala Asn Arg
35 40 45
Gly His Asn Val Thr Val Val Thr Ser Phe Pro Gln Lys Lys Pro Val
50 55 60
Pro Asn Leu Tyr Glu Ile Asp Val Ser Gly Ala Lys Gly Leu Ala Thr
65 70 75 80
Asn Ser Ile His Phe Glu Arg Leu Gln Thr Ile Ile Gln Asp Val Lys
85 90 95
Ser Asn Phe Lys Asn Met Val Arg Leu Ser Arg Thr Tyr Cys Glu Ile
100 105 110
Met Phe Ser Asp Pro Arg Val Leu Asn Ile Arg Asp Lys Lys Phe Asp
115 120 125
Leu Val Ile Asn Ala Val Phe Gly Ser Asp Cys Asp Ala Gly Phe Ala
130 135 140
Trp Lys Ser Gln Ala Pro Leu Ile Ser Ile Leu Asn Ala Arg His Thr
145 150 155 160
Pro Trp Ala Leu His Arg Met Gly Asn Pro Ser Asn Pro Ala Tyr Met
165 170 175
Pro Val Ile His Ser Arg Phe Pro Val Lys Met Asn Phe Phe Gln Arg
180 185 190
Met Ile Asn Thr Gly Trp His Leu Tyr Phe Leu Tyr Met Tyr Phe Tyr
195 200 205
Tyr Gly Asn Gly Glu Asp Ala Asn Lys Met Ala Arg Lys Phe Phe Gly
210 215 220
Asn Asp Met Pro Asp Ile Asn Glu Met Val Phe Asn Thr Ser Leu Leu
225 230 235 240
Phe Val Asn Thr His Phe Ser Val Asp Met Pro Tyr Pro Leu Val Pro
245 250 255
Asn Cys Ile Glu Ile Gly Gly Ile His Val Lys Glu Pro Gln Pro Leu
260 265 270
Pro Leu Glu Ile Gln Lys Phe Met Asp Glu Ala Glu His Gly Val Ile
275 280 285
Phe Phe Thr Leu Gly Ser Met Val Arg Thr Ser Thr Phe Pro Asn Gln
290 295 300
Thr Ile Gln Ala Phe Lys Glu Ala Phe Ala Glu Leu Pro Gln Arg Val
305 310 315 320
Leu Trp Lys Phe Glu Asn Glu Asn Glu Asp Met Pro Ser Asn Val Leu
325 330 335
Ile Arg Lys Trp Phe Pro Gln Asn Asp Ile Phe Gly His Lys Asn Ile
340 345 350
Lys Ala Phe Ile Ser His Gly Gly Asn Ser Gly Ala Leu Glu Ala Val
355 360 365
His Phe Gly Val Pro Ile Ile Gly Ile Pro Leu Phe Tyr Asp Gln Tyr
370 375 380
Arg Asn Ile Leu Ser Phe Val Lys Glu Gly Val Ala Val Leu Leu Asp
385 390 395 400
Val Asn Asp Leu Thr Lys Asp Asn Ile Leu Ser Ser Val Arg Thr Val
405 410 415
Val Asn Asp Lys Ser Tyr Ser Glu Arg Met Lys Ala Leu Ser Gln Leu
420 425 430
Phe Arg Asp Arg Pro Met Ser Pro Leu Asp Thr Ala Val Tyr Trp Thr
435 440 445
Glu Tyr Val Ile Arg His Arg Gly Ala His His Leu Lys Thr Ala Gly
450 455 460
Ala Phe Leu His Trp Tyr Gln Tyr Leu Leu Leu Asp Val Ile Thr Phe
465 470 475 480
Leu Leu Val Thr Phe Cys Ala Phe Cys Phe Ile Val Lys Tyr Ile Cys
485 490 495
Lys Ala Leu Ile His His Tyr Trp Ser Ser Ser Lys Ser Glu Lys Leu
500 505 510
Lys Lys Asn
515
<210> 20
<211> 1548
<212> DNA
<213> 胭脂虫
<400> 20
atggaattca gattgttgat cttggctttg ttctctgttt tgatgtctac ttctaacggt 60
gctgaaatct tggctttgtt cccaatccac ggtatctcta actacaacgt tgctgaagct 120
ttgttgaaga ctttggctaa cagaggtcac aacgttactg ttgttacttc tttcccacaa 180
aagaagccag ttccaaactt gtacgaaatc gacgtttctg gtgctaaggg tttggctact 240
aactctatcc acttcgaaag attgcaaact atcatccaag acgttaagtc taacttcaag 300
aacatggtta gattgtctag aacttactgt gaaatcatgt tctctgaccc aagagttttg 360
aacatcagag acaagaagtt cgacttggtt atcaacgctg ttttcggttc tgactgtgac 420
gctggtttcg cttggaagtc tcaagctcca ttgatctcta tcttgaacgc tagacacact 480
ccatgggctt tgcacagaat gggtaaccca tctaacccag cttacatgcc agttatccac 540
tctagattcc cagttaagat gaacttcttc caaagaatga tcaacactgg ttggcacttg 600
tacttcttgt acatgtactt ctactacggt aacggtgaag acgctaacaa gatggctaga 660
aagttcttcg gtaacgacat gccagacatc aacgaaatgg ttttcaacac ttctttgttg 720
ttcgttaaca ctcacttctc tgttgacatg ccatacccat tggttccaaa ctgtatcgaa 780
atcggtggta tccacgttaa ggaaccacaa ccattgccat tggaaatcca aaagttcatg 840
gacgaagctg aacacggtgt tatcttcttc actttgggtt ctatggttag aacttctact 900
ttcccaaacc aaactatcca agctttcaag gaagctttcg ctgaattgcc acaaagagtt 960
ttgtggaagt tcgaaaacga aaacgaagac atgccatcta acgttttgat cagaaagtgg 1020
ttcccacaaa acgacatctt cggtcacaag aacatcaagg ctttcatctc tcacggtggt 1080
aactctggtg ctttggaagc tgttcacttc ggtgttccaa tcatcggtat cccattgttc 1140
tacgaccaat acagaaacat cttgtctttc gttaaggaag gtgttgctgt tttgttggac 1200
gttaacgact tgactaagga caacatcttg tcttctgtta gaactgttgt taacgacaag 1260
tcttactctg aaagaatgaa ggctttgtct caattgttca gagacagacc aatgtctcca 1320
ttggacactg ctgtttactg gactgaatac gttatcagac acagaggtgc tcaccacttg 1380
aagactgctg gtgctttctt gcactggtac caatacttgt tgttggacgt tatcactttc 1440
ttgttggtta ctttctgtgc tttctgtttc atcgttaagt acatctgtaa ggctttgatc 1500
caccactact ggtcttcttc taagtctgaa aagttgaaga agaactag 1548
<210> 21
<211> 504
<212> PRT
<213> 胭脂虫
<400> 21
Met Thr Leu Leu Arg Asp Leu Leu Leu Leu Tyr Ile Asn Ser Leu Leu
1 5 10 15
Phe Ile Asn Pro Ser Ile Gly Glu Asn Ile Leu Val Phe Leu Pro Thr
20 25 30
Lys Thr Tyr Ser His Phe Lys Pro Leu Glu Pro Leu Phe Gln Glu Leu
35 40 45
Ala Met Arg Gly His Asn Val Thr Val Phe Ser Gly Phe Ser Leu Thr
50 55 60
Lys Asn Ile Ser Asn Tyr Ser Ser Ile Val Phe Ser Ala Glu Ile Glu
65 70 75 80
Phe Val Asn Ile Gly Met Gly Asn Leu Arg Lys Gln Ser Arg Ile Tyr
85 90 95
Asn Trp Ile Tyr Val His Asn Glu Leu Gln Asn Tyr Phe Thr Gln Leu
100 105 110
Ile Ser Asp Asn Gln Leu Gln Glu Leu Leu Ser Asn Lys Asp Thr Gln
115 120 125
Phe Asp Leu Ile Phe Ile Glu Leu Tyr His Val Asp Gly Val Phe Ala
130 135 140
Leu Ser His Arg Phe Asn Cys Pro Ile Ile Gly Leu Ser Phe Gln Pro
145 150 155 160
Val Leu Pro Ile Tyr Asn Trp Leu Ile Gly Asn Pro Thr Thr Phe Ser
165 170 175
Tyr Ile Pro His Val Tyr Leu Pro Phe Thr Asp Ile Met Ser Phe Trp
180 185 190
Lys Arg Ile Ile Asn Ala Val Phe Ser Ile Phe Thr Ala Ala Phe Tyr
195 200 205
Asn Phe Val Ser Thr Lys Gly Tyr Gln Lys His Val Asp Leu Leu Leu
210 215 220
Arg Gln Thr Glu Ser Pro Lys Leu Asn Ile Glu Glu Leu Ser Glu Ser
225 230 235 240
Leu Ser Leu Ile Leu Ala Glu Phe His Phe Ser Ser Ala Tyr Thr Arg
245 250 255
Pro Asn Leu Pro Asn Val Ile Asp Ile Ala Gly Ile His Ile Gln Ser
260 265 270
Pro Lys Pro Leu Pro Gln Asp Leu Leu Asp Phe Leu Asp Gln Ser Glu
275 280 285
His Gly Val Ile Tyr Val Ser Leu Gly Thr Leu Ile Asp Pro Ile His
290 295 300
Thr Asp His Leu Gly Leu Asn Leu Ile Asn Val Phe Arg Lys Leu Arg
305 310 315 320
Gln Arg Val Ile Trp Lys Trp Lys Lys Glu Phe Phe His Asp Val Pro
325 330 335
Lys Asn Val Leu Ile Gly Glu Trp Phe Pro Gln Ile Asp Ile Leu Asn
340 345 350
His Pro Arg Cys Lys Leu Phe Ile Ser His Gly Gly Tyr His Ser Met
355 360 365
Leu Glu Ser Ile Tyr Ser Ser Val Pro Ile Leu Gly Ile Pro Phe Phe
370 375 380
Thr Asp Gln His His Asn Thr Ala Ile Ile Glu Lys Leu Lys Ile Gly
385 390 395 400
Lys Lys Ala Ser Thr Glu Ala Ser Glu Glu Asp Leu Leu Thr Ala Val
405 410 415
Lys Glu Leu Leu Ser Asn Glu Thr Phe Lys Arg Asn Ser Gln His Gln
420 425 430
Ser Ser Ile Phe Arg Asp Arg Pro Met Ser Pro Met Asp Thr Ala Ile
435 440 445
Tyr Trp Thr Glu Tyr Ile Leu Arg Tyr Lys Gly Ala Ser His Met Lys
450 455 460
Ser Ala Val Ile Asp Leu Tyr Trp Phe Gln Tyr Ile Leu Leu Asp Ile
465 470 475 480
Ile Leu Phe Tyr Ser Leu Ile Val Leu Ile Leu Leu Cys Ile Leu Arg
485 490 495
Ile Phe Phe Arg Met Leu Thr Lys
500
<210> 22
<211> 1515
<212> DNA
<213> 胭脂虫
<400> 22
atgactttgt tgagagactt gttgttgttg tacatcaact ctttgttgtt catcaaccca 60
tctatcggtg aaaacatctt ggttttcttg ccaactaaga cttactctca cttcaagcca 120
ttggaaccat tgttccaaga attggctatg agaggtcaca acgttactgt tttctctggt 180
ttctctttga ctaagaacat ctctaactac tcttctatcg ttttctctgc tgaaatcgaa 240
ttcgttaaca tcggtatggg taacttgaga aagcaatcta gaatctacaa ctggatctac 300
gttcacaacg aattgcaaaa ctacttcact caattgatct ctgacaacca attgcaagaa 360
ttgttgtcta acaaggacac tcaattcgac ttgatcttca tcgaattgta ccacgttgac 420
ggtgttttcg ctttgtctca cagattcaac tgtccaatca tcggtttgtc tttccaacca 480
gttttgccaa tctacaactg gttgatcggt aacccaacta ctttctctta catcccacac 540
gtttacttgc cattcactga catcatgtct ttctggaaga gaatcatcaa cgctgttttc 600
tctatcttca ctgctgcttt ctacaacttc gtttctacta agggttacca aaagcacgtt 660
gacttgttgt tgagacaaac tgaatctcca aagttgaaca tcgaagaatt gtctgaatct 720
ttgtctttga tcttggctga attccacttc tcttctgctt acactagacc aaacttgcca 780
aacgttatcg acatcgctgg tatccacatc caatctccaa agccattgcc acaagacttg 840
ttggacttct tggaccaatc tgaacacggt gttatctacg tttctttggg tactttgatc 900
gacccaatcc acactgacca cttgggtttg aacttgatca acgttttcag aaagttgaga 960
caaagagtta tctggaagtg gaagaaggaa ttcttccacg acgttccaaa gaacgttttg 1020
atcggtgaat ggttcccaca aatcgacatc ttgaaccacc caagatgtaa gttgttcatc 1080
tctcacggtg gttaccactc tatgttggaa tctatctact cttctgttcc aatcttgggt 1140
atcccattct tcactgacca acaccacaac actgctatca tcgaaaagtt gaagatcggt 1200
aagaaggctt ctactgaagc ttctgaagaa gacttgttga ctgctgttaa ggaattgttg 1260
tctaacgaaa ctttcaagag aaactctcaa caccaatctt ctatcttcag agacagacca 1320
atgtctccaa tggacactgc tatctactgg actgaataca tcttgagata caagggtgct 1380
tctcacatga agtctgctgt tatcgacttg tactggttcc aatacatctt gttggacatc 1440
atcttgttct actctttgat cgttttgatc ttgttgtgta tcttgagaat cttcttcaga 1500
atgttgacta agtag 1515
<210> 23
<211> 526
<212> PRT
<213> 胭脂虫
<400> 23
Met Ile Phe Phe Tyr Phe Leu Thr Leu Thr Ser Phe Ile Ser Val Ala
1 5 10 15
Phe Ser Tyr Asn Ile Leu Gly Val Phe Pro Phe Gln Ala Lys Ser His
20 25 30
Phe Gly Phe Ile Asp Pro Leu Leu Val Arg Leu Ala Glu Leu Gly His
35 40 45
Asn Val Thr Ile Tyr Asp Pro Tyr Pro Lys Ser Glu Lys Leu Pro Asn
50 55 60
Tyr Asn Glu Ile Asp Val Ser Glu Cys Phe Val Phe Asn Thr Leu Tyr
65 70 75 80
Glu Glu Ile Asp Thr Phe Ile Lys Thr Ala Ala Ser Pro Phe Ser Ser
85 90 95
Leu Trp Tyr Ser Phe Glu Glu Thr Leu Ala Val Phe Gln Lys Glu Asn
100 105 110
Phe Asp Lys Cys Ala Pro Leu Arg Glu Leu Leu Asn Ser Thr Val Lys
115 120 125
Tyr Asp Leu Leu Ile Thr Glu Thr Phe Leu Thr Asp Ile Thr Leu Leu
130 135 140
Phe Val Asn Lys Phe Lys Ile Pro Phe Ile Thr Ser Thr Pro Asn Val
145 150 155 160
Pro Phe Pro Trp Leu Ala Asp Arg Met Gly Asn Pro Leu Asn Pro Ser
165 170 175
Tyr Ile Pro Asn Leu Phe Ser Asp Tyr Pro Phe Asp Lys Met Thr Phe
180 185 190
Phe Asn Arg Leu Trp Asn Thr Leu Phe Tyr Val Met Ala Leu Gly Gly
195 200 205
His Asn Ala Ile Ile Leu Lys Asn Glu Glu Lys Ile Asn Lys Tyr Tyr
210 215 220
Phe Gly Ser Ser Val Pro Ser Leu Tyr Asn Ile Ala Arg Glu Thr Ser
225 230 235 240
Ile Met Leu Ile Asn Ala His Glu Thr Leu Asn Pro Val Ile Pro Leu
245 250 255
Val Pro Gly Met Ile Pro Val Ser Gly Ile His Ile Lys Gln Pro Ala
260 265 270
Ala Leu Pro Gln Asn Ile Glu Lys Phe Ile Asn Glu Ser Thr His Gly
275 280 285
Val Val Tyr Phe Cys Met Gly Ser Leu Leu Arg Gly Glu Thr Phe Pro
290 295 300
Ala Glu Lys Arg Asp Ala Phe Leu Tyr Ala Phe Ser Lys Ile Pro Gln
305 310 315 320
Arg Val Leu Trp Lys Trp Glu Gly Glu Val Leu Pro Gly Lys Ser Glu
325 330 335
Asn Ile Met Thr Ser Lys Trp Met Pro Gln Arg Asp Ile Leu Ala His
340 345 350
Pro Asn Val Lys Leu Phe Ile Ser His Gly Gly Leu Leu Gly Thr Ser
355 360 365
Glu Ala Val Tyr Glu Gly Val Pro Val Ile Gly Ile Pro Ile Phe Gly
370 375 380
Asp Gln Arg Thr Asn Ile Lys Ala Leu Glu Ala Asn Gly Ala Gly Glu
385 390 395 400
Leu Leu Asp Tyr Asn Asp Ile Ser Gly Glu Val Val Leu Glu Lys Ile
405 410 415
Gln Arg Leu Ile Asn Asp Pro Lys Tyr Lys Glu Ser Ala Arg Gln Leu
420 425 430
Ser Ile Arg Tyr Lys Asp Arg Pro Met Ser Pro Leu Asp Thr Ala Val
435 440 445
Tyr Trp Thr Glu Tyr Val Ile Arg His Lys Gly Ala Pro His Leu Lys
450 455 460
Thr Ala Ala Val Asp Met Pro Trp Tyr Gln Tyr Leu Leu Leu Asp Val
465 470 475 480
Ile Ala Phe Leu Ile Phe Ile Leu Val Ser Val Ile Leu Ile Ile Tyr
485 490 495
Tyr Gly Val Lys Ile Ser Leu Arg Tyr Leu Cys Ala Leu Ile Phe Gly
500 505 510
Asn Ser Ser Ser Leu Lys Pro Thr Lys Lys Val Lys Asp Asn
515 520 525
<210> 24
<211> 1581
<212> DNA
<213> 胭脂虫
<400> 24
atgatcttct tctacttctt gactttgact tctttcatct ctgttgcttt ctcttacaac 60
atcttgggtg ttttcccatt ccaagctaag tctcacttcg gtttcatcga cccattgttg 120
gttagattgg ctgaattggg tcacaacgtt actatctacg acccataccc aaagtctgaa 180
aagttgccaa actacaacga aatcgacgtt tctgaatgtt tcgttttcaa cactttgtac 240
gaagaaatcg acactttcat caagactgct gcttctccat tctcttcttt gtggtactct 300
ttcgaagaaa ctttggctgt tttccaaaag gaaaacttcg acaagtgtgc tccattgaga 360
gaattgttga actctactgt taagtacgac ttgttgatca ctgaaacttt cttgactgac 420
atcactttgt tgttcgttaa caagttcaag atcccattca tcacttctac tccaaacgtt 480
ccattcccat ggttggctga cagaatgggt aacccattga acccatctta catcccaaac 540
ttgttctctg actacccatt cgacaagatg actttcttca acagattgtg gaacactttg 600
ttctacgtta tggctttggg tggtcacaac gctatcatct tgaagaacga agaaaagatc 660
aacaagtact acttcggttc ttctgttcca tctttgtaca acatcgctag agaaacttct 720
atcatgttga tcaacgctca cgaaactttg aacccagtta tcccattggt tccaggtatg 780
atcccagttt ctggtatcca catcaagcaa ccagctgctt tgccacaaaa catcgaaaag 840
ttcatcaacg aatctactca cggtgttgtt tacttctgta tgggttcttt gttgagaggt 900
gaaactttcc cagctgaaaa gagagacgct ttcttgtacg ctttctctaa gatcccacaa 960
agagttttgt ggaagtggga aggtgaagtt ttgccaggta agtctgaaaa catcatgact 1020
tctaagtgga tgccacaaag agacatcttg gctcacccaa acgttaagtt gttcatctct 1080
cacggtggtt tgttgggtac ttctgaagct gtttacgaag gtgttccagt tatcggtatc 1140
ccaatcttcg gtgaccaaag aactaacatc aaggctttgg aagctaacgg tgctggtgaa 1200
ttgttggact acaacgacat ctctggtgaa gttgttttgg aaaagatcca aagattgatc 1260
aacgacccaa agtacaagga atctgctaga caattgtcta tcagatacaa ggacagacca 1320
atgtctccat tggacactgc tgtttactgg actgaatacg ttatcagaca caagggtgct 1380
ccacacttga agactgctgc tgttgacatg ccatggtacc aatacttgtt gttggacgtt 1440
atcgctttct tgatcttcat cttggtttct gttatcttga tcatctacta cggtgttaag 1500
atctctttga gatacttgtg tgctttgatc ttcggtaact cttcttcttt gaagccaact 1560
aagaaggtta aggacaacta g 1581
<210> 25
<211> 484
<212> PRT
<213> 拟南芥
<400> 25
Met Asn Arg Glu Val Ser Glu Arg Ile His Ile Leu Phe Phe Pro Phe
1 5 10 15
Met Ala Gln Gly His Met Ile Pro Ile Leu Asp Met Ala Lys Leu Phe
20 25 30
Ser Arg Arg Gly Ala Lys Ser Thr Leu Leu Thr Thr Pro Ile Asn Ala
35 40 45
Lys Ile Phe Glu Lys Pro Ile Glu Ala Phe Lys Asn Gln Asn Pro Asp
50 55 60
Leu Glu Ile Gly Ile Lys Ile Phe Asn Phe Pro Cys Val Glu Leu Gly
65 70 75 80
Leu Pro Glu Gly Cys Glu Asn Ala Asp Phe Ile Asn Ser Tyr Gln Lys
85 90 95
Ser Asp Ser Gly Asp Leu Phe Leu Lys Phe Leu Phe Ser Thr Lys Tyr
100 105 110
Met Lys Gln Gln Leu Glu Ser Phe Ile Glu Thr Thr Lys Pro Ser Ala
115 120 125
Leu Val Ala Asp Met Phe Phe Pro Trp Ala Thr Glu Ser Ala Glu Lys
130 135 140
Leu Gly Val Pro Arg Leu Val Phe His Gly Thr Ser Phe Phe Ser Leu
145 150 155 160
Cys Cys Ser Tyr Asn Met Arg Ile His Lys Pro His Lys Lys Val Ala
165 170 175
Thr Ser Ser Thr Pro Phe Val Ile Pro Gly Leu Pro Gly Asp Ile Val
180 185 190
Ile Thr Glu Asp Gln Ala Asn Val Ala Lys Glu Glu Thr Pro Met Gly
195 200 205
Lys Phe Met Lys Glu Val Arg Glu Ser Glu Thr Asn Ser Phe Gly Val
210 215 220
Leu Val Asn Ser Phe Tyr Glu Leu Glu Ser Ala Tyr Ala Asp Phe Tyr
225 230 235 240
Arg Ser Phe Val Ala Lys Arg Ala Trp His Ile Gly Pro Leu Ser Leu
245 250 255
Ser Asn Arg Glu Leu Gly Glu Lys Ala Arg Arg Gly Lys Lys Ala Asn
260 265 270
Ile Asp Glu Gln Glu Cys Leu Lys Trp Leu Asp Ser Lys Thr Pro Gly
275 280 285
Ser Val Val Tyr Leu Ser Phe Gly Ser Gly Thr Asn Phe Thr Asn Asp
290 295 300
Gln Leu Leu Glu Ile Ala Phe Gly Leu Glu Gly Ser Gly Gln Ser Phe
305 310 315 320
Ile Trp Val Val Arg Lys Asn Glu Asn Gln Gly Asp Asn Glu Glu Trp
325 330 335
Leu Pro Glu Gly Phe Lys Glu Arg Thr Thr Gly Lys Gly Leu Ile Ile
340 345 350
Pro Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Lys Ala Ile Gly
355 360 365
Gly Phe Val Thr His Cys Gly Trp Asn Ser Ala Ile Glu Gly Ile Ala
370 375 380
Ala Gly Leu Pro Met Val Thr Trp Pro Met Gly Ala Glu Gln Phe Tyr
385 390 395 400
Asn Glu Lys Leu Leu Thr Lys Val Leu Arg Ile Gly Val Asn Val Gly
405 410 415
Ala Thr Glu Leu Val Lys Lys Gly Lys Leu Ile Ser Arg Ala Gln Val
420 425 430
Glu Lys Ala Val Arg Glu Val Ile Gly Gly Glu Lys Ala Glu Glu Arg
435 440 445
Arg Leu Trp Ala Lys Lys Leu Gly Glu Met Ala Lys Ala Ala Val Glu
450 455 460
Glu Gly Gly Ser Ser Tyr Asn Asp Val Asn Lys Phe Met Glu Glu Leu
465 470 475 480
Asn Gly Arg Lys
<210> 26
<211> 1455
<212> DNA
<213> 拟南芥
<400> 26
atgaacagag aagtttctga aagaatccac atcttgttct tcccattcat ggctcaaggt 60
cacatgatcc caatcttgga catggctaag ttgttctcta gaagaggtgc taagtctact 120
ttgttgacta ctccaatcaa cgctaagatc ttcgaaaagc caatcgaagc tttcaagaac 180
caaaacccag acttggaaat cggtatcaag atcttcaact tcccatgtgt tgaattgggt 240
ttgccagaag gttgtgaaaa cgctgacttc atcaactctt accaaaagtc tgactctggt 300
gacttgttct tgaagttctt gttctctact aagtacatga agcaacaatt ggaatctttc 360
atcgaaacta ctaagccatc tgctttggtt gctgacatgt tcttcccatg ggctactgaa 420
tctgctgaaa agttgggtgt tccaagattg gttttccacg gtacttcttt cttctctttg 480
tgttgttctt acaacatgag aatccacaag ccacacaaga aggttgctac ttcttctact 540
ccattcgtta tcccaggttt gccaggtgac atcgttatca ctgaagacca agctaacgtt 600
gctaaggaag aaactccaat gggtaagttc atgaaggaag ttagagaatc tgaaactaac 660
tctttcggtg ttttggttaa ctctttctac gaattggaat ctgcttacgc tgacttctac 720
agatctttcg ttgctaagag agcttggcac atcggtccat tgtctttgtc taacagagaa 780
ttgggtgaaa aggctagaag aggtaagaag gctaacatcg acgaacaaga atgtttgaag 840
tggttggact ctaagactcc aggttctgtt gtttacttgt ctttcggttc tggtactaac 900
ttcactaacg accaattgtt ggaaatcgct ttcggtttgg aaggttctgg tcaatctttc 960
atctgggttg ttagaaagaa cgaaaaccaa ggtgacaacg aagaatggtt gccagaaggt 1020
ttcaaggaaa gaactactgg taagggtttg atcatcccag gttgggctcc acaagttttg 1080
atcttggacc acaaggctat cggtggtttc gttactcact gtggttggaa ctctgctatc 1140
gaaggtatcg ctgctggttt gccaatggtt acttggccaa tgggtgctga acaattctac 1200
aacgaaaagt tgttgactaa ggttttgaga atcggtgtta acgttggtgc tactgaattg 1260
gttaagaagg gtaagttgat ctctagagct caagttgaaa aggctgttag agaagttatc 1320
ggtggtgaaa aggctgaaga aagaagattg tgggctaaga agttgggtga aatggctaag 1380
gctgctgttg aagaaggtgg ttcttcttac aacgacgtta acaagttcat ggaagaattg 1440
aacggtagaa agtag 1455
<210> 27
<211> 455
<212> PRT
<213> 拟南芥
<400> 27
Met Glu Lys Ser Asn Gly Leu Arg Val Ile Leu Phe Pro Leu Pro Leu
1 5 10 15
Gln Gly Cys Ile Asn Pro Met Ile Gln Leu Ala Lys Ile Leu His Ser
20 25 30
Arg Gly Phe Ser Ile Thr Val Ile His Thr Cys Phe Asn Ala Pro Lys
35 40 45
Ala Ser Ser His Pro Leu Phe Thr Phe Leu Glu Ile Pro Asp Gly Leu
50 55 60
Ser Glu Thr Glu Lys Arg Thr Asn Asn Thr Lys Leu Leu Leu Thr Leu
65 70 75 80
Leu Asn Arg Asn Cys Glu Ser Pro Phe Arg Glu Cys Leu Ser Lys Leu
85 90 95
Leu Gln Ser Ala Asp Ser Glu Thr Gly Glu Glu Lys Gln Arg Ile Ser
100 105 110
Cys Leu Ile Ala Asp Ser Gly Trp Met Phe Thr Gln Pro Ile Ala Gln
115 120 125
Ser Leu Lys Leu Pro Ile Leu Val Leu Ser Val Phe Thr Val Ser Phe
130 135 140
Phe Arg Cys Gln Phe Val Leu Pro Lys Leu Arg Arg Glu Val Tyr Leu
145 150 155 160
Pro Leu Gln Asp Ser Glu Gln Glu Asp Leu Val Gln Glu Phe Pro Pro
165 170 175
Leu Arg Lys Lys Asp Ile Val Arg Ile Leu Asp Val Glu Thr Asp Ile
180 185 190
Leu Asp Pro Phe Leu Asp Lys Val Leu Gln Met Thr Lys Ala Ser Ser
195 200 205
Gly Leu Ile Phe Met Ser Cys Glu Glu Leu Asp His Asp Ser Val Ser
210 215 220
Gln Ala Arg Glu Asp Phe Lys Ile Pro Ile Phe Gly Ile Gly Pro Ser
225 230 235 240
His Ser His Phe Pro Ala Thr Ser Ser Ser Leu Ser Thr Pro Asp Glu
245 250 255
Thr Cys Ile Pro Trp Leu Asp Lys Gln Glu Asp Lys Ser Val Ile Tyr
260 265 270
Val Ser Tyr Gly Ser Ile Val Thr Ile Ser Glu Ser Asp Leu Ile Glu
275 280 285
Ile Ala Trp Gly Leu Arg Asn Ser Asp Gln Pro Phe Leu Leu Val Val
290 295 300
Arg Val Gly Ser Val Arg Gly Arg Glu Trp Ile Glu Thr Ile Pro Glu
305 310 315 320
Glu Ile Met Glu Lys Leu Asn Glu Lys Gly Lys Ile Val Lys Trp Ala
325 330 335
Pro Gln Gln Asp Val Leu Lys His Arg Ala Ile Gly Gly Phe Leu Thr
340 345 350
His Asn Gly Trp Ser Ser Thr Val Glu Ser Val Cys Glu Ala Val Pro
355 360 365
Met Ile Cys Leu Pro Phe Arg Trp Asp Gln Met Leu Asn Ala Arg Phe
370 375 380
Val Ser Asp Val Trp Met Val Gly Ile Asn Leu Glu Asp Arg Val Glu
385 390 395 400
Arg Asn Glu Ile Glu Gly Ala Ile Arg Arg Leu Leu Val Glu Pro Glu
405 410 415
Gly Glu Ala Ile Arg Glu Arg Ile Glu His Leu Lys Glu Lys Val Gly
420 425 430
Arg Ser Phe Gln Gln Asn Gly Ser Ala Tyr Gln Ser Leu Gln Asn Leu
435 440 445
Ile Asp Tyr Ile Ser Ser Phe
450 455
<210> 28
<211> 1368
<212> DNA
<213> 拟南芥
<400> 28
atggaaaagt ctaacggttt gagagttatc ttgttcccat tgccattgca aggttgtatc 60
aacccaatga tccaattggc taagatcttg cactctagag gtttctctat cactgttatc 120
cacacttgtt tcaacgctcc aaaggcttct tctcacccat tgttcacttt cttggaaatc 180
ccagacggtt tgtctgaaac tgaaaagaga actaacaaca ctaagttgtt gttgactttg 240
ttgaacagaa actgtgaatc tccattcaga gaatgtttgt ctaagttgtt gcaatctgct 300
gactctgaaa ctggtgaaga aaagcaaaga atctcttgtt tgatcgctga ctctggttgg 360
atgttcactc aaccaatcgc tcaatctttg aagttgccaa tcttggtttt gtctgttttc 420
actgtttctt tcttcagatg tcaattcgtt ttgccaaagt tgagaagaga agtttacttg 480
ccattgcaag actctgaaca agaagacttg gttcaagaat tcccaccatt gagaaagaag 540
gacatcgtta gaatcttgga cgttgaaact gacatcttgg acccattctt ggacaaggtt 600
ttgcaaatga ctaaggcttc ttctggtttg atcttcatgt cttgtgaaga attggaccac 660
gactctgttt ctcaagctag agaagacttc aagatcccaa tcttcggtat cggtccatct 720
cactctcact tcccagctac ttcttcttct ttgtctactc cagacgaaac ttgtatccca 780
tggttggaca agcaagaaga caagtctgtt atctacgttt cttacggttc tatcgttact 840
atctctgaat ctgacttgat cgaaatcgct tggggtttga gaaactctga ccaaccattc 900
ttgttggttg ttagagttgg ttctgttaga ggtagagaat ggatcgaaac tatcccagaa 960
gaaatcatgg aaaagttgaa cgaaaagggt aagatcgtta agtgggctcc acaacaagac 1020
gttttgaagc acagagctat cggtggtttc ttgactcaca acggttggtc ttctactgtt 1080
gaatctgttt gtgaagctgt tccaatgatc tgtttgccat tcagatggga ccaaatgttg 1140
aacgctagat tcgtttctga cgtttggatg gttggtatca acttggaaga cagagttgaa 1200
agaaacgaaa tcgaaggtgc tatcagaaga ttgttggttg aaccagaagg tgaagctatc 1260
agagaaagaa tcgaacactt gaaggaaaag gttggtagat ctttccaaca aaacggttct 1320
gcttaccaat ctttgcaaaa cttgatcgac tacatctctt ctttctag 1368
<210> 29
<211> 481
<212> PRT
<213> 拟南芥
<400> 29
Met Ser Ser Asp Pro His Arg Lys Leu His Val Val Phe Phe Pro Phe
1 5 10 15
Met Ala Tyr Gly His Met Ile Pro Thr Leu Asp Met Ala Lys Leu Phe
20 25 30
Ser Ser Arg Gly Ala Lys Ser Thr Ile Leu Thr Thr Pro Leu Asn Ser
35 40 45
Lys Ile Phe Gln Lys Pro Ile Glu Arg Phe Lys Asn Leu Asn Pro Ser
50 55 60
Phe Glu Ile Asp Ile Gln Ile Phe Asp Phe Pro Cys Val Asp Leu Gly
65 70 75 80
Leu Pro Glu Gly Cys Glu Asn Val Asp Phe Phe Thr Ser Asn Asn Asn
85 90 95
Asp Asp Arg Gln Tyr Leu Thr Leu Lys Phe Phe Lys Ser Thr Arg Phe
100 105 110
Phe Lys Asp Gln Leu Glu Lys Leu Leu Glu Thr Thr Arg Pro Asp Cys
115 120 125
Leu Ile Ala Asp Met Phe Phe Pro Trp Ala Thr Glu Ala Ala Glu Lys
130 135 140
Phe Asn Val Pro Arg Leu Val Phe His Gly Thr Gly Tyr Phe Ser Leu
145 150 155 160
Cys Ser Glu Tyr Cys Ile Arg Val His Asn Pro Gln Asn Ile Val Ala
165 170 175
Ser Arg Tyr Glu Pro Phe Val Ile Pro Asp Leu Pro Gly Asn Ile Val
180 185 190
Ile Thr Gln Glu Gln Ile Ala Asp Arg Asp Glu Glu Ser Glu Met Gly
195 200 205
Lys Phe Met Ile Glu Val Lys Glu Ser Asp Val Lys Ser Ser Gly Val
210 215 220
Ile Val Asn Ser Phe Tyr Glu Leu Glu Pro Asp Tyr Ala Asp Phe Tyr
225 230 235 240
Lys Ser Val Val Leu Lys Arg Ala Trp His Ile Gly Pro Leu Ser Val
245 250 255
Tyr Asn Arg Gly Phe Glu Glu Lys Ala Glu Arg Gly Lys Lys Ala Ser
260 265 270
Ile Asn Glu Val Glu Cys Leu Lys Trp Leu Asp Ser Lys Lys Pro Asp
275 280 285
Ser Val Ile Tyr Ile Ser Phe Gly Ser Val Ala Cys Phe Lys Asn Glu
290 295 300
Gln Leu Phe Glu Ile Ala Ala Gly Leu Glu Thr Ser Gly Ala Asn Phe
305 310 315 320
Ile Trp Val Val Arg Lys Asn Ile Gly Ile Glu Lys Glu Glu Trp Leu
325 330 335
Pro Glu Gly Phe Glu Glu Arg Val Lys Gly Lys Gly Met Ile Ile Arg
340 345 350
Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Gln Ala Thr Cys Gly
355 360 365
Phe Val Thr His Cys Gly Trp Asn Ser Leu Leu Glu Gly Val Ala Ala
370 375 380
Gly Leu Pro Met Val Thr Trp Pro Val Ala Ala Glu Gln Phe Tyr Asn
385 390 395 400
Glu Lys Leu Val Thr Gln Val Leu Arg Thr Gly Val Ser Val Gly Ala
405 410 415
Lys Lys Asn Val Arg Thr Thr Gly Asp Phe Ile Ser Arg Glu Lys Val
420 425 430
Val Lys Ala Val Arg Glu Val Leu Val Gly Glu Glu Ala Asp Glu Arg
435 440 445
Arg Glu Arg Ala Lys Lys Leu Ala Glu Met Ala Lys Ala Ala Val Glu
450 455 460
Gly Gly Ser Ser Phe Asn Asp Leu Asn Ser Phe Ile Glu Glu Phe Thr
465 470 475 480
Ser
<210> 30
<211> 1446
<212> DNA
<213> 拟南芥
<400> 30
atgtcttctg acccacacag aaagttgcac gttgttttct tcccattcat ggcttacggt 60
cacatgatcc caactttgga catggctaag ttgttctctt ctagaggtgc taagtctact 120
atcttgacta ctccattgaa ctctaagatc ttccaaaagc caatcgaaag attcaagaac 180
ttgaacccat ctttcgaaat cgacatccaa atcttcgact tcccatgtgt tgacttgggt 240
ttgccagaag gttgtgaaaa cgttgacttc ttcacttcta acaacaacga cgacagacaa 300
tacttgactt tgaagttctt caagtctact agattcttca aggaccaatt ggaaaagttg 360
ttggaaacta ctagaccaga ctgtttgatc gctgacatgt tcttcccatg ggctactgaa 420
gctgctgaaa agttcaacgt tccaagattg gttttccacg gtactggtta cttctctttg 480
tgttctgaat actgtatcag agttcacaac ccacaaaaca tcgttgcttc tagatacgaa 540
ccattcgtta tcccagactt gccaggtaac atcgttatca ctcaagaaca aatcgctgac 600
agagacgaag aatctgaaat gggtaagttc atgatcgaag ttaaggaatc tgacgttaag 660
tcttctggtg ttatcgttaa ctctttctac gaattggaac cagactacgc tgacttctac 720
aagtctgttg ttttgaagag agcttggcac atcggtccat tgtctgttta caacagaggt 780
ttcgaagaaa aggctgaaag aggtaagaag gcttctatca acgaagttga atgtttgaag 840
tggttggact ctaagaagcc agactctgtt atctacatct ctttcggttc tgttgcttgt 900
ttcaagaacg aacaattgtt cgaaatcgct gctggtttgg aaacttctgg tgctaacttc 960
atctgggttg ttagaaagaa catcggtatc gaaaaggaag aatggttgcc agaaggtttc 1020
gaagaaagag ttaagggtaa gggtatgatc atcagaggtt gggctccaca agttttgatc 1080
ttggaccacc aagctacttg tggtttcgtt actcactgtg gttggaactc tttgttggaa 1140
ggtgttgctg ctggtttgcc aatggttact tggccagttg ctgctgaaca attctacaac 1200
gaaaagttgg ttactcaagt tttgagaact ggtgtttctg ttggtgctaa gaagaacgtt 1260
agaactactg gtgacttcat ctctagagaa aaggttgtta aggctgttag agaagttttg 1320
gttggtgaag aagctgacga aagaagagaa agagctaaga agttggctga aatggctaag 1380
gctgctgttg aaggtggttc ttctttcaac gacttgaact ctttcatcga agaattcact 1440
tcttag 1446
<210> 31
<211> 474
<212> PRT
<213> 甜叶菊
<400> 31
Met Ser Thr Ser Glu Leu Val Phe Ile Pro Ser Pro Gly Ala Gly His
1 5 10 15
Leu Pro Pro Thr Val Glu Leu Ala Lys Leu Leu Leu His Arg Asp Gln
20 25 30
Arg Leu Ser Val Thr Ile Ile Val Met Asn Leu Trp Leu Gly Pro Lys
35 40 45
His Asn Thr Glu Ala Arg Pro Cys Val Pro Ser Leu Arg Phe Val Asp
50 55 60
Ile Pro Cys Asp Glu Ser Thr Met Ala Leu Ile Ser Pro Asn Thr Phe
65 70 75 80
Ile Ser Ala Phe Val Glu His His Lys Pro Arg Val Arg Asp Ile Val
85 90 95
Arg Gly Ile Ile Glu Ser Asp Ser Val Arg Leu Ala Gly Phe Val Leu
100 105 110
Asp Met Phe Cys Met Pro Met Ser Asp Val Ala Asn Glu Phe Gly Val
115 120 125
Pro Ser Tyr Asn Tyr Phe Thr Ser Gly Ala Ala Thr Leu Gly Leu Met
130 135 140
Phe His Leu Gln Trp Lys Arg Asp His Glu Gly Tyr Asp Ala Thr Glu
145 150 155 160
Leu Lys Asn Ser Asp Thr Glu Leu Ser Val Pro Ser Tyr Val Asn Pro
165 170 175
Val Pro Ala Lys Val Leu Pro Glu Val Val Leu Asp Lys Glu Gly Gly
180 185 190
Ser Lys Met Phe Leu Asp Leu Ala Glu Arg Ile Arg Glu Ser Lys Gly
195 200 205
Ile Ile Val Asn Ser Cys Gln Ala Ile Glu Arg His Ala Leu Glu Tyr
210 215 220
Leu Ser Ser Asn Asn Asn Gly Ile Pro Pro Val Phe Pro Val Gly Pro
225 230 235 240
Ile Leu Asn Leu Glu Asn Lys Lys Asp Asp Ala Lys Thr Asp Glu Ile
245 250 255
Met Arg Trp Leu Asn Glu Gln Pro Glu Ser Ser Val Val Phe Leu Cys
260 265 270
Phe Gly Ser Met Gly Ser Phe Asn Glu Lys Gln Val Lys Glu Ile Ala
275 280 285
Val Ala Ile Glu Arg Ser Gly His Arg Phe Leu Trp Ser Leu Arg Arg
290 295 300
Pro Thr Pro Lys Glu Lys Ile Glu Phe Pro Lys Glu Tyr Glu Asn Leu
305 310 315 320
Glu Glu Val Leu Pro Glu Gly Phe Leu Lys Arg Thr Ser Ser Ile Gly
325 330 335
Lys Val Ile Gly Trp Ala Pro Gln Met Ala Val Leu Ser His Pro Ser
340 345 350
Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Thr Leu Glu Ser
355 360 365
Met Trp Cys Gly Val Pro Met Ala Ala Trp Pro Leu Tyr Ala Glu Gln
370 375 380
Thr Leu Asn Ala Phe Leu Leu Val Val Glu Leu Gly Leu Ala Ala Glu
385 390 395 400
Ile Arg Met Asp Tyr Arg Thr Asp Thr Lys Ala Gly Tyr Asp Gly Gly
405 410 415
Met Glu Val Thr Val Glu Glu Ile Glu Asp Gly Ile Arg Lys Leu Met
420 425 430
Ser Asp Gly Glu Ile Arg Asn Lys Val Lys Asp Val Lys Glu Lys Ser
435 440 445
Arg Ala Ala Val Val Glu Gly Gly Ser Ser Tyr Ala Ser Ile Gly Lys
450 455 460
Phe Ile Glu His Val Ser Asn Val Thr Ile
465 470
<210> 32
<211> 1425
<212> DNA
<213> 甜叶菊
<400> 32
atgtctactt ctgaattggt tttcatccca tctccaggtg ctggtcactt gccaccaact 60
gttgaattgg ctaagttgtt gttgcacaga gaccaaagat tgtctgttac tatcatcgtt 120
atgaacttgt ggttgggtcc aaagcacaac actgaagcta gaccatgtgt tccatctttg 180
agattcgttg acatcccatg tgacgaatct actatggctt tgatctctcc aaacactttc 240
atctctgctt tcgttgaaca ccacaagcca agagttagag acatcgttag aggtatcatc 300
gaatctgact ctgttagatt ggctggtttc gttttggaca tgttctgtat gccaatgtct 360
gacgttgcta acgaattcgg tgttccatct tacaactact tcacttctgg tgctgctact 420
ttgggtttga tgttccactt gcaatggaag agagaccacg aaggttacga cgctactgaa 480
ttgaagaact ctgacactga attgtctgtt ccatcttacg ttaacccagt tccagctaag 540
gttttgccag aagttgtttt ggacaaggaa ggtggttcta agatgttctt ggacttggct 600
gaaagaatca gagaatctaa gggtatcatc gttaactctt gtcaagctat cgaaagacac 660
gctttggaat acttgtcttc taacaacaac ggtatcccac cagttttccc agttggtcca 720
atcttgaact tggaaaacaa gaaggacgac gctaagactg acgaaatcat gagatggttg 780
aacgaacaac cagaatcttc tgttgttttc ttgtgtttcg gttctatggg ttctttcaac 840
gaaaagcaag ttaaggaaat cgctgttgct atcgaaagat ctggtcacag attcttgtgg 900
tctttgagaa gaccaactcc aaaggaaaag atcgaattcc caaaggaata cgaaaacttg 960
gaagaagttt tgccagaagg tttcttgaag agaacttctt ctatcggtaa ggttatcggt 1020
tgggctccac aaatggctgt tttgtctcac ccatctgttg gtggtttcgt ttctcactgt 1080
ggttggaact ctactttgga atctatgtgg tgtggtgttc caatggctgc ttggccattg 1140
tacgctgaac aaactttgaa cgctttcttg ttggttgttg aattgggttt ggctgctgaa 1200
atcagaatgg actacagaac tgacactaag gctggttacg acggtggtat ggaagttact 1260
gttgaagaaa tcgaagacgg tatcagaaag ttgatgtctg acggtgaaat cagaaacaag 1320
gttaaggacg ttaaggaaaa gtctagagct gctgttgttg aaggtggttc ttcttacgct 1380
tctatcggta agttcatcga acacgtttct aacgttacta tctag 1425
<210> 33
<211> 478
<212> PRT
<213> 水稻
<400> 33
Met Lys Gln Thr Val Val Leu Tyr Pro Gly Gly Gly Val Gly His Val
1 5 10 15
Val Pro Met Leu Glu Leu Ala Lys Val Phe Val Lys His Gly His Asp
20 25 30
Val Thr Met Val Leu Leu Glu Pro Pro Phe Lys Ser Ser Asp Ser Gly
35 40 45
Ala Leu Ala Val Glu Arg Leu Val Ala Ser Asn Pro Ser Val Ser Phe
50 55 60
His Val Leu Pro Pro Leu Pro Ala Pro Asp Phe Ala Ser Phe Gly Lys
65 70 75 80
His Pro Phe Leu Leu Val Ile Gln Leu Leu Arg Gln Tyr Asn Glu Arg
85 90 95
Leu Glu Ser Phe Leu Leu Ser Ile Pro Arg Gln Arg Leu His Ser Leu
100 105 110
Val Ile Asp Met Phe Cys Val Asp Ala Ile Asp Val Cys Ala Lys Leu
115 120 125
Gly Val Pro Val Tyr Thr Phe Phe Ala Ser Gly Val Ser Val Leu Ser
130 135 140
Val Leu Thr Gln Leu Pro Pro Phe Leu Ala Gly Arg Glu Thr Gly Leu
145 150 155 160
Lys Glu Leu Gly Asp Thr Pro Leu Asp Phe Leu Gly Val Ser Pro Met
165 170 175
Pro Ala Ser His Leu Val Lys Glu Leu Leu Glu His Pro Glu Asp Glu
180 185 190
Leu Cys Lys Ala Met Val Asn Arg Trp Glu Arg Asn Thr Glu Thr Met
195 200 205
Gly Val Leu Val Asn Ser Phe Glu Ser Leu Glu Ser Arg Ala Ala Gln
210 215 220
Ala Leu Arg Asp Asp Pro Leu Cys Val Pro Gly Lys Val Leu Pro Pro
225 230 235 240
Ile Tyr Cys Val Gly Pro Leu Val Gly Gly Gly Ala Glu Glu Ala Ala
245 250 255
Glu Arg His Glu Cys Leu Val Trp Leu Asp Ala Gln Pro Glu His Ser
260 265 270
Val Val Phe Leu Cys Phe Gly Ser Lys Gly Val Phe Ser Ala Glu Gln
275 280 285
Leu Lys Glu Ile Ala Val Gly Leu Glu Asn Ser Arg Gln Arg Phe Met
290 295 300
Trp Val Val Arg Thr Pro Pro Thr Thr Thr Glu Gly Leu Lys Lys Tyr
305 310 315 320
Phe Glu Gln Arg Ala Ala Pro Asp Leu Asp Ala Leu Phe Pro Asp Gly
325 330 335
Phe Val Glu Arg Thr Lys Asp Arg Gly Phe Ile Val Thr Thr Trp Ala
340 345 350
Pro Gln Val Asp Val Leu Arg His Arg Ala Thr Gly Ala Phe Val Thr
355 360 365
His Cys Gly Trp Asn Ser Ala Leu Glu Gly Ile Thr Ala Gly Val Pro
370 375 380
Met Leu Cys Trp Pro Gln Tyr Ala Glu Gln Lys Met Asn Lys Val Phe
385 390 395 400
Met Thr Ala Glu Met Gly Val Gly Val Glu Leu Asp Gly Tyr Asn Ser
405 410 415
Asp Phe Val Lys Ala Glu Glu Leu Glu Ala Lys Val Arg Leu Val Met
420 425 430
Glu Ser Glu Glu Gly Lys Gln Leu Arg Ala Arg Ser Ala Ala Arg Lys
435 440 445
Lys Glu Ala Glu Ala Ala Leu Glu Glu Gly Gly Ser Ser His Ala Ala
450 455 460
Phe Val Gln Phe Leu Ser Asp Val Glu Asn Leu Val Gln Asn
465 470 475
<210> 34
<211> 1437
<212> DNA
<213> 水稻
<400> 34
atgaagcaaa ctgttgtttt gtacccaggt ggtggtgttg gtcacgttgt tccaatgttg 60
gaattggcta aggttttcgt taagcacggt cacgacgtta ctatggtttt gttggaacca 120
ccattcaagt cttctgactc tggtgctttg gctgttgaaa gattggttgc ttctaaccca 180
tctgtttctt tccacgtttt gccaccattg ccagctccag acttcgcttc tttcggtaag 240
cacccattct tgttggttat ccaattgttg agacaataca acgaaagatt ggaatctttc 300
ttgttgtcta tcccaagaca aagattgcac tctttggtta tcgacatgtt ctgtgttgac 360
gctatcgacg tttgtgctaa gttgggtgtt ccagtttaca ctttcttcgc ttctggtgtt 420
tctgttttgt ctgttttgac tcaattgcca ccattcttgg ctggtagaga aactggtttg 480
aaggaattgg gtgacactcc attggacttc ttgggtgttt ctccaatgcc agcttctcac 540
ttggttaagg aattgttgga acacccagaa gacgaattgt gtaaggctat ggttaacaga 600
tgggaaagaa acactgaaac tatgggtgtt ttggttaact ctttcgaatc tttggaatct 660
agagctgctc aagctttgag agacgaccca ttgtgtgttc caggtaaggt tttgccacca 720
atctactgtg ttggtccatt ggttggtggt ggtgctgaag aagctgctga aagacacgaa 780
tgtttggttt ggttggacgc tcaaccagaa cactctgttg ttttcttgtg tttcggttct 840
aagggtgttt tctctgctga acaattgaag gaaatcgctg ttggtttgga aaactctaga 900
caaagattca tgtgggttgt tagaactcca ccaactacta ctgaaggttt gaagaagtac 960
ttcgaacaaa gagctgctcc agacttggac gctttgttcc cagacggttt cgttgaaaga 1020
actaaggaca gaggtttcat cgttactact tgggctccac aagttgacgt tttgagacac 1080
agagctactg gtgctttcgt tactcactgt ggttggaact ctgctttgga aggtatcact 1140
gctggtgttc caatgttgtg ttggccacaa tacgctgaac aaaagatgaa caaggttttc 1200
atgactgctg aaatgggtgt tggtgttgaa ttggacggtt acaactctga cttcgttaag 1260
gctgaagaat tggaagctaa ggttagattg gttatggaat ctgaagaagg taagcaattg 1320
agagctagat ctgctgctag aaagaaggaa gctgaagctg ctttggaaga aggtggttct 1380
tctcacgctg ctttcgttca attcttgtct gacgttgaaa acttggttca aaactag 1437
<210> 35
<211> 530
<212> PRT
<213> 智人
<400> 35
Met Ala Arg Ala Gly Trp Thr Ser Pro Val Pro Leu Cys Val Cys Leu
1 5 10 15
Leu Leu Thr Cys Gly Phe Ala Glu Ala Gly Lys Leu Leu Val Val Pro
20 25 30
Met Asp Gly Ser His Trp Phe Thr Met Gln Ser Val Val Glu Lys Leu
35 40 45
Ile Leu Arg Gly His Glu Val Val Val Val Met Pro Glu Val Ser Trp
50 55 60
Gln Leu Glu Arg Ser Leu Asn Cys Thr Val Lys Thr Tyr Ser Thr Ser
65 70 75 80
Tyr Thr Leu Glu Asp Gln Asn Arg Glu Phe Met Val Phe Ala His Ala
85 90 95
Gln Trp Lys Ala Gln Ala Gln Ser Ile Phe Ser Leu Leu Met Ser Ser
100 105 110
Ser Ser Gly Phe Leu Asp Leu Phe Phe Ser His Cys Arg Ser Leu Phe
115 120 125
Asn Asp Arg Lys Leu Val Glu Tyr Leu Lys Glu Ser Ser Phe Asp Ala
130 135 140
Val Phe Leu Asp Pro Phe Asp Thr Cys Gly Leu Ile Val Ala Lys Tyr
145 150 155 160
Phe Ser Leu Pro Ser Val Val Phe Thr Arg Gly Ile Phe Cys His His
165 170 175
Leu Glu Glu Gly Ala Gln Cys Pro Ala Pro Leu Ser Tyr Val Pro Asn
180 185 190
Asp Leu Leu Gly Phe Ser Asp Ala Met Thr Phe Lys Glu Arg Val Trp
195 200 205
Asn His Ile Val His Leu Glu Asp His Leu Phe Cys Gln Tyr Leu Phe
210 215 220
Arg Asn Ala Leu Glu Ile Ala Ser Glu Ile Leu Gln Thr Pro Val Thr
225 230 235 240
Ala Tyr Asp Leu Tyr Ser His Thr Ser Ile Trp Leu Leu Arg Thr Asp
245 250 255
Phe Val Leu Asp Tyr Pro Lys Pro Val Met Pro Asn Met Ile Phe Ile
260 265 270
Gly Gly Ile Asn Cys His Gln Gly Lys Pro Leu Pro Met Glu Phe Glu
275 280 285
Ala Tyr Ile Asn Ala Ser Gly Glu His Gly Ile Val Val Phe Ser Leu
290 295 300
Gly Ser Met Val Ser Glu Ile Pro Glu Lys Lys Ala Met Ala Ile Ala
305 310 315 320
Asp Ala Leu Gly Lys Ile Pro Gln Thr Val Leu Trp Arg Tyr Thr Gly
325 330 335
Thr Arg Pro Ser Asn Leu Ala Asn Asn Thr Ile Leu Val Lys Trp Leu
340 345 350
Pro Gln Asn Asp Leu Leu Gly His Pro Met Thr Arg Ala Phe Ile Thr
355 360 365
His Ala Gly Ser His Gly Val Tyr Glu Ser Ile Cys Asn Gly Val Pro
370 375 380
Met Val Met Met Pro Leu Phe Gly Asp Gln Met Asp Asn Ala Lys Arg
385 390 395 400
Met Glu Thr Lys Gly Ala Gly Val Thr Leu Asn Val Leu Glu Met Thr
405 410 415
Ser Glu Asp Leu Glu Asn Ala Leu Lys Ala Val Ile Asn Asp Lys Ser
420 425 430
Tyr Lys Glu Asn Ile Met Arg Leu Ser Ser Leu His Lys Asp Arg Pro
435 440 445
Val Glu Pro Leu Asp Leu Ala Val Phe Trp Val Glu Phe Val Met Arg
450 455 460
His Lys Gly Ala Pro His Leu Arg Pro Ala Ala His Asp Leu Thr Trp
465 470 475 480
Tyr Gln Tyr His Ser Leu Asp Val Ile Gly Phe Leu Leu Ala Val Val
485 490 495
Leu Thr Val Ala Phe Ile Thr Phe Lys Cys Cys Ala Tyr Gly Tyr Arg
500 505 510
Lys Cys Leu Gly Lys Lys Gly Arg Val Lys Lys Ala His Lys Ser Lys
515 520 525
Thr His
530
<210> 36
<211> 1590
<212> DNA
<213> 智人
<400> 36
atggctagag ctggttggac ttctccagtt ccattgtgtg tttgtttgtt gttgacttgt 60
ggtttcgctg aagctggtaa gttgttggtt gttccaatgg acggttctca ctggttcact 120
atgcaatctg ttgttgaaaa gttgatcttg agaggtcacg aagttgttgt tgttatgcca 180
gaagtttctt ggcaattgga aagatctttg aactgtactg ttaagactta ctctacttct 240
tacactttgg aagaccaaaa cagagaattc atggttttcg ctcacgctca atggaaggct 300
caagctcaat ctatcttctc tttgttgatg tcttcttctt ctggtttctt ggacttgttc 360
ttctctcact gtagatcttt gttcaacgac agaaagttgg ttgaatactt gaaggaatct 420
tctttcgacg ctgttttctt ggacccattc gacacttgtg gtttgatcgt tgctaagtac 480
ttctctttgc catctgttgt tttcactaga ggtatcttct gtcaccactt ggaagaaggt 540
gctcaatgtc cagctccatt gtcttacgtt ccaaacgact tgttgggttt ctctgacgct 600
atgactttca aggaaagagt ttggaaccac atcgttcact tggaagacca cttgttctgt 660
caatacttgt tcagaaacgc tttggaaatc gcttctgaaa tcttgcaaac tccagttact 720
gcttacgact tgtactctca cacttctatc tggttgttga gaactgactt cgttttggac 780
tacccaaagc cagttatgcc aaacatgatc ttcatcggtg gtatcaactg tcaccaaggt 840
aagccattgc caatggaatt cgaagcttac atcaacgctt ctggtgaaca cggtatcgtt 900
gttttctctt tgggttctat ggtttctgaa atcccagaaa agaaggctat ggctatcgct 960
gacgctttgg gtaagatccc acaaactgtt ttgtggagat acactggtac tagaccatct 1020
aacttggcta acaacactat cttggttaag tggttgccac aaaacgactt gttgggtcac 1080
ccaatgacta gagctttcat cactcacgct ggttctcacg gtgtttacga atctatctgt 1140
aacggtgttc caatggttat gatgccattg ttcggtgacc aaatggacaa cgctaagaga 1200
atggaaacta agggtgctgg tgttactttg aacgttttgg aaatgacttc tgaagacttg 1260
gaaaacgctt tgaaggctgt tatcaacgac aagtcttaca aggaaaacat catgagattg 1320
tcttctttgc acaaggacag accagttgaa ccattggact tggctgtttt ctgggttgaa 1380
ttcgttatga gacacaaggg tgctccacac ttgagaccag ctgctcacga cttgacttgg 1440
taccaatacc actctttgga cgttatcggt ttcttgttgg ctgttgtttt gactgttgct 1500
ttcatcactt tcaagtgttg tgcttacggt tacagaaagt gtttgggtaa gaagggtaga 1560
gttaagaagg ctcacaagtc taagactcac 1590
<210> 37
<211> 530
<212> PRT
<213> 智人
<400> 37
Met Ala Cys Thr Gly Trp Thr Ser Pro Leu Pro Leu Cys Val Cys Leu
1 5 10 15
Leu Leu Thr Cys Gly Phe Ala Glu Ala Gly Lys Leu Leu Val Val Pro
20 25 30
Met Asp Gly Ser His Trp Phe Thr Met Arg Ser Val Val Glu Lys Leu
35 40 45
Ile Leu Arg Gly His Glu Val Val Val Val Met Pro Glu Val Ser Trp
50 55 60
Gln Leu Gly Arg Ser Leu Asn Cys Thr Val Lys Thr Tyr Ser Thr Ser
65 70 75 80
Tyr Thr Leu Glu Asp Leu Asp Arg Glu Phe Lys Ala Phe Ala His Ala
85 90 95
Gln Trp Lys Ala Gln Val Arg Ser Ile Tyr Ser Leu Leu Met Gly Ser
100 105 110
Tyr Asn Asp Ile Phe Asp Leu Phe Phe Ser Asn Cys Arg Ser Leu Phe
115 120 125
Lys Asp Lys Lys Leu Val Glu Tyr Leu Lys Glu Ser Ser Phe Asp Ala
130 135 140
Val Phe Leu Asp Pro Phe Asp Asn Cys Gly Leu Ile Val Ala Lys Tyr
145 150 155 160
Phe Ser Leu Pro Ser Val Val Phe Ala Arg Gly Ile Leu Cys His Tyr
165 170 175
Leu Glu Glu Gly Ala Gln Cys Pro Ala Pro Leu Ser Tyr Val Pro Arg
180 185 190
Ile Leu Leu Gly Phe Ser Asp Ala Met Thr Phe Lys Glu Arg Val Arg
195 200 205
Asn His Ile Met His Leu Glu Glu His Leu Leu Cys His Arg Phe Phe
210 215 220
Lys Asn Ala Leu Glu Ile Ala Ser Glu Ile Leu Gln Thr Pro Val Thr
225 230 235 240
Glu Tyr Asp Leu Tyr Ser His Thr Ser Ile Trp Leu Leu Arg Thr Asp
245 250 255
Phe Val Leu Asp Tyr Pro Lys Pro Val Met Pro Asn Met Ile Phe Ile
260 265 270
Gly Gly Ile Asn Cys His Gln Gly Lys Pro Leu Pro Met Glu Phe Glu
275 280 285
Ala Tyr Ile Asn Ala Ser Gly Glu His Gly Ile Val Val Phe Ser Leu
290 295 300
Gly Ser Met Val Ser Glu Ile Pro Glu Lys Lys Ala Met Ala Ile Ala
305 310 315 320
Asp Ala Leu Gly Lys Ile Pro Gln Thr Val Leu Trp Arg Tyr Thr Gly
325 330 335
Thr Arg Pro Ser Asn Leu Ala Asn Asn Thr Ile Leu Val Lys Trp Leu
340 345 350
Pro Gln Asn Asp Leu Leu Gly His Pro Met Thr Arg Ala Phe Ile Thr
355 360 365
His Ala Gly Ser His Gly Val Tyr Glu Ser Ile Cys Asn Gly Val Pro
370 375 380
Met Val Met Met Pro Leu Phe Gly Asp Gln Met Asp Asn Ala Lys Arg
385 390 395 400
Met Glu Thr Lys Gly Ala Gly Val Thr Leu Asn Val Leu Glu Met Thr
405 410 415
Ser Glu Asp Leu Glu Asn Ala Leu Lys Ala Val Ile Asn Asp Lys Ser
420 425 430
Tyr Lys Glu Asn Ile Met Arg Leu Ser Ser Leu His Lys Asp Arg Pro
435 440 445
Val Glu Pro Leu Asp Leu Ala Val Phe Trp Val Glu Phe Val Met Arg
450 455 460
His Lys Gly Ala Pro His Leu Arg Pro Ala Ala His Asp Leu Thr Trp
465 470 475 480
Tyr Gln Tyr His Ser Leu Asp Val Ile Gly Phe Leu Leu Ala Val Val
485 490 495
Leu Thr Val Ala Phe Ile Thr Phe Lys Cys Cys Ala Tyr Gly Tyr Arg
500 505 510
Lys Cys Leu Gly Lys Lys Gly Arg Val Lys Lys Ala His Lys Ser Lys
515 520 525
Thr His
530
<210> 38
<211> 1590
<212> DNA
<213> 智人
<400> 38
atggcttgta ctggttggac ttctccattg ccattgtgtg tttgtttgtt gttgacttgt 60
ggtttcgctg aagctggtaa gttgttggtt gttccaatgg acggttctca ctggttcact 120
atgagatctg ttgttgaaaa gttgatcttg agaggtcacg aagttgttgt tgttatgcca 180
gaagtttctt ggcaattggg tagatctttg aactgtactg ttaagactta ctctacttct 240
tacactttgg aagacttgga cagagaattc aaggctttcg ctcacgctca atggaaggct 300
caagttagat ctatctactc tttgttgatg ggttcttaca acgacatctt cgacttgttc 360
ttctctaact gtagatcttt gttcaaggac aagaagttgg ttgaatactt gaaggaatct 420
tctttcgacg ctgttttctt ggacccattc gacaactgtg gtttgatcgt tgctaagtac 480
ttctctttgc catctgttgt tttcgctaga ggtatcttgt gtcactactt ggaagaaggt 540
gctcaatgtc cagctccatt gtcttacgtt ccaagaatct tgttgggttt ctctgacgct 600
atgactttca aggaaagagt tagaaaccac atcatgcact tggaagaaca cttgttgtgt 660
cacagattct tcaagaacgc tttggaaatc gcttctgaaa tcttgcaaac tccagttact 720
gaatacgact tgtactctca cacttctatc tggttgttga gaactgactt cgttttggac 780
tacccaaagc cagttatgcc aaacatgatc ttcatcggtg gtatcaactg tcaccaaggt 840
aagccattgc caatggaatt cgaagcttac atcaacgctt ctggtgaaca cggtatcgtt 900
gttttctctt tgggttctat ggtttctgaa atcccagaaa agaaggctat ggctatcgct 960
gacgctttgg gtaagatccc acaaactgtt ttgtggagat acactggtac tagaccatct 1020
aacttggcta acaacactat cttggttaag tggttgccac aaaacgactt gttgggtcac 1080
ccaatgacta gagctttcat cactcacgct ggttctcacg gtgtttacga atctatctgt 1140
aacggtgttc caatggttat gatgccattg ttcggtgacc aaatggacaa cgctaagaga 1200
atggaaacta agggtgctgg tgttactttg aacgttttgg aaatgacttc tgaagacttg 1260
gaaaacgctt tgaaggctgt tatcaacgac aagtcttaca aggaaaacat catgagattg 1320
tcttctttgc acaaggacag accagttgaa ccattggact tggctgtttt ctgggttgaa 1380
ttcgttatga gacacaaggg tgctccacac ttgagaccag ctgctcacga cttgacttgg 1440
taccaatacc actctttgga cgttatcggt ttcttgttgg ctgttgtttt gactgttgct 1500
ttcatcactt tcaagtgttg tgcttacggt tacagaaagt gtttgggtaa gaagggtaga 1560
gttaagaagg ctcacaagtc taagactcac 1590
<210> 39
<211> 529
<212> PRT
<213> 智人
<400> 39
Met Ser Val Lys Trp Thr Ser Val Ile Leu Leu Ile Gln Leu Ser Phe
1 5 10 15
Cys Phe Ser Ser Gly Asn Cys Gly Lys Val Leu Val Trp Ala Ala Glu
20 25 30
Tyr Ser His Trp Met Asn Ile Lys Thr Ile Leu Asp Glu Leu Ile Gln
35 40 45
Arg Gly His Glu Val Thr Val Leu Ala Ser Ser Ala Ser Ile Leu Phe
50 55 60
Asp Pro Asn Asn Ser Ser Ala Leu Lys Ile Glu Ile Tyr Pro Thr Ser
65 70 75 80
Leu Thr Lys Thr Glu Leu Glu Asn Phe Ile Met Gln Gln Ile Lys Arg
85 90 95
Trp Ser Asp Leu Pro Lys Asp Thr Phe Trp Leu Tyr Phe Ser Gln Val
100 105 110
Gln Glu Ile Met Ser Ile Phe Gly Asp Ile Thr Arg Lys Phe Cys Lys
115 120 125
Asp Val Val Ser Asn Lys Lys Phe Met Lys Lys Val Gln Glu Ser Arg
130 135 140
Phe Asp Val Ile Phe Ala Asp Ala Ile Phe Pro Cys Ser Glu Leu Leu
145 150 155 160
Ala Glu Leu Phe Asn Ile Pro Phe Val Tyr Ser Leu Ser Phe Ser Pro
165 170 175
Gly Tyr Thr Phe Glu Lys His Ser Gly Gly Phe Ile Phe Pro Pro Ser
180 185 190
Tyr Val Pro Val Val Met Ser Glu Leu Thr Asp Gln Met Thr Phe Met
195 200 205
Glu Arg Val Lys Asn Met Ile Tyr Val Leu Tyr Phe Asp Phe Trp Phe
210 215 220
Glu Ile Phe Asp Met Lys Lys Trp Asp Gln Phe Tyr Ser Glu Val Leu
225 230 235 240
Gly Arg Pro Thr Thr Leu Ser Glu Thr Met Gly Lys Ala Asp Val Trp
245 250 255
Leu Ile Arg Asn Ser Trp Asn Phe Gln Phe Pro Tyr Pro Leu Leu Pro
260 265 270
Asn Val Asp Phe Val Gly Gly Leu His Cys Lys Pro Ala Lys Pro Leu
275 280 285
Pro Lys Glu Met Glu Asp Phe Val Gln Ser Ser Gly Glu Asn Gly Val
290 295 300
Val Val Phe Ser Leu Gly Ser Met Val Ser Asn Met Thr Glu Glu Arg
305 310 315 320
Ala Asn Val Ile Ala Ser Ala Leu Ala Gln Ile Pro Gln Lys Val Leu
325 330 335
Trp Arg Phe Asp Gly Asn Lys Pro Asp Thr Leu Gly Leu Asn Thr Arg
340 345 350
Leu Tyr Lys Trp Ile Pro Gln Asn Asp Leu Leu Gly His Pro Lys Thr
355 360 365
Arg Ala Phe Ile Thr His Gly Gly Ala Asn Gly Ile Tyr Glu Ala Ile
370 375 380
Tyr His Gly Ile Pro Met Val Gly Ile Pro Leu Phe Ala Asp Gln Pro
385 390 395 400
Asp Asn Ile Ala His Met Lys Ala Arg Gly Ala Ala Val Arg Val Asp
405 410 415
Phe Asn Thr Met Ser Ser Thr Asp Leu Leu Asn Ala Leu Lys Arg Val
420 425 430
Ile Asn Asp Pro Ser Tyr Lys Glu Asn Val Met Lys Leu Ser Arg Ile
435 440 445
Gln His Asp Gln Pro Val Lys Pro Leu Asp Arg Ala Val Phe Trp Ile
450 455 460
Glu Phe Val Met Arg His Lys Gly Ala Lys His Leu Arg Val Ala Ala
465 470 475 480
His Asp Leu Thr Trp Phe Gln Tyr His Ser Leu Asp Val Ile Gly Phe
485 490 495
Leu Leu Val Cys Val Ala Thr Val Ile Phe Ile Val Thr Lys Cys Cys
500 505 510
Leu Phe Cys Phe Trp Lys Phe Ala Arg Lys Ala Lys Lys Gly Lys Asn
515 520 525
Asp
<210> 40
<211> 1587
<212> DNA
<213> 智人
<400> 40
atgtctgtta agtggacttc tgttatcttg ttgatccaat tgtctttctg tttctcttct 60
ggtaactgtg gtaaggtttt ggtttgggct gctgaatact ctcactggat gaacatcaag 120
actatcttgg acgaattgat ccaaagaggt cacgaagtta ctgttttggc ttcttctgct 180
tctatcttgt tcgacccaaa caactcttct gctttgaaga tcgaaatcta cccaacttct 240
ttgactaaga ctgaattgga aaacttcatc atgcaacaaa tcaagagatg gtctgacttg 300
ccaaaggaca ctttctggtt gtacttctct caagttcaag aaatcatgtc tatcttcggt 360
gacatcacta gaaagttctg taaggacgtt gtttctaaca agaagttcat gaagaaggtt 420
caagaatcta gattcgacgt tatcttcgct gacgctatct tcccatgttc tgaattgttg 480
gctgaattgt tcaacatccc attcgtttac tctttgtctt tctctccagg ttacactttc 540
gaaaagcact ctggtggttt catcttccca ccatcttacg ttccagttgt tatgtctgaa 600
ttgactgacc aaatgacttt catggaaaga gttaagaaca tgatctacgt tttgtacttc 660
gacttctggt tcgaaatctt cgacatgaag aagtgggacc aattctactc tgaagttttg 720
ggtagaccaa ctactttgtc tgaaactatg ggtaaggctg acgtttggtt gatcagaaac 780
tcttggaact tccaattccc atacccattg ttgccaaacg ttgacttcgt tggtggtttg 840
cactgtaagc cagctaagcc attgccaaag gaaatggaag acttcgttca atcttctggt 900
gaaaacggtg ttgttgtttt ctctttgggt tctatggttt ctaacatgac tgaagaaaga 960
gctaacgtta tcgcttctgc tttggctcaa atcccacaaa aggttttgtg gagattcgac 1020
ggtaacaagc cagacacttt gggtttgaac actagattgt acaagtggat cccacaaaac 1080
gacttgttgg gtcacccaaa gactagagct ttcatcactc acggtggtgc taacggtatc 1140
tacgaagcta tctaccacgg tatcccaatg gttggtatcc cattgttcgc tgaccaacca 1200
gacaacatcg ctcacatgaa ggctagaggt gctgctgtta gagttgactt caacactatg 1260
tcttctactg acttgttgaa cgctttgaag agagttatca acgacccatc ttacaaggaa 1320
aacgttatga agttgtctag aatccaacac gaccaaccag ttaagccatt ggacagagct 1380
gttttctgga tcgaattcgt tatgagacac aagggtgcta agcacttgag agttgctgct 1440
cacgacttga cttggttcca ataccactct ttggacgtta tcggtttctt gttggtttgt 1500
gttgctactg ttatcttcat cgttactaag tgttgtttgt tctgtttctg gaagttcgct 1560
agaaaggcta agaagggtaa gaacgac 1587
<210> 41
<400> 41
000
<210> 42
<400> 42
000
<210> 43
<400> 43
000
<210> 44
<400> 44
000
<210> 45
<211> 296
<212> PRT
<213> 拟南芥
<400> 45
Met Phe Asp Phe Asn Lys Tyr Met Asp Ser Lys Ala Met Thr Val Asn
1 5 10 15
Glu Ala Leu Asn Lys Ala Ile Pro Leu Arg Tyr Pro Gln Lys Ile Tyr
20 25 30
Glu Ser Met Arg Tyr Ser Leu Leu Ala Gly Gly Lys Arg Val Arg Pro
35 40 45
Val Leu Cys Ile Ala Ala Cys Glu Leu Val Gly Gly Thr Glu Glu Leu
50 55 60
Ala Ile Pro Thr Ala Cys Ala Ile Glu Met Ile His Thr Met Ser Leu
65 70 75 80
Met His Asp Asp Leu Pro Cys Ile Asp Asn Asp Asp Leu Arg Arg Gly
85 90 95
Lys Pro Thr Asn His Lys Ile Phe Gly Glu Asp Thr Ala Val Thr Ala
100 105 110
Gly Asn Ala Leu His Ser Tyr Ala Phe Glu His Ile Ala Val Ser Thr
115 120 125
Ser Lys Thr Val Gly Ala Asp Arg Ile Leu Arg Met Val Ser Glu Leu
130 135 140
Gly Arg Ala Thr Gly Ser Glu Gly Val Met Gly Gly Gln Met Val Asp
145 150 155 160
Ile Ala Ser Glu Gly Asp Pro Ser Ile Asp Leu Gln Thr Leu Glu Trp
165 170 175
Ile His Ile His Lys Thr Ala Met Leu Leu Glu Cys Ser Val Val Cys
180 185 190
Gly Ala Ile Ile Gly Gly Ala Ser Glu Ile Val Ile Glu Arg Ala Arg
195 200 205
Arg Tyr Ala Arg Cys Val Gly Leu Leu Phe Gln Val Val Asp Asp Ile
210 215 220
Leu Asp Val Thr Lys Ser Ser Asp Glu Leu Gly Lys Thr Ala Gly Lys
225 230 235 240
Asp Leu Ile Ser Asp Lys Ala Thr Tyr Pro Lys Leu Met Gly Leu Glu
245 250 255
Lys Ala Lys Glu Phe Ser Asp Glu Leu Leu Asn Arg Ala Lys Gly Glu
260 265 270
Leu Ser Cys Phe Asp Pro Val Lys Ala Ala Pro Leu Leu Gly Leu Ala
275 280 285
Asp Tyr Val Ala Phe Arg Gln Asn
290 295
<210> 46
<211> 891
<212> DNA
<213> 拟南芥
<400> 46
atgttcgact tcaacaagta catggactct aaggctatga ctgttaacga agctttgaac 60
aaggctatcc cattgagata cccacaaaag atctacgaat ctatgagata ctctttgttg 120
gctggtggta agagagttag accagttttg tgtatcgctg cttgtgaatt ggttggtggt 180
actgaagaat tggctatccc aactgcttgt gctatcgaaa tgatccacac tatgtctttg 240
atgcacgacg acttgccatg tatcgacaac gacgacttga gaagaggtaa gccaactaac 300
cacaagatct tcggtgaaga cactgctgtt actgctggta acgctttgca ctcttacgct 360
ttcgaacaca tcgctgtttc tacttctaag actgttggtg ctgacagaat cttgagaatg 420
gtttctgaat tgggtagagc tactggttct gaaggtgtta tgggtggtca aatggttgac 480
atcgcttctg aaggtgaccc atctatcgac ttgcaaactt tggaatggat ccacatccac 540
aagactgcta tgttgttgga atgttctgtt gtttgtggtg ctatcatcgg tggtgcttct 600
gaaatcgtta tcgaaagagc tagaagatac gctagatgtg ttggtttgtt gttccaagtt 660
gttgacgaca tcttggacgt tactaagtct tctgacgaat tgggtaagac tgctggtaag 720
gacttgatct ctgacaaggc tacttaccca aagttgatgg gtttggaaaa ggctaaggaa 780
ttctctgacg aattgttgaa cagagctaag ggtgaattgt cttgtttcga cccagttaag 840
gctgctccat tgttgggttt ggctgactac gttgctttca gacaaaacta g 891
<210> 47
<211> 720
<212> PRT
<213> 大麻
<400> 47
Met Gly Lys Asn Tyr Lys Ser Leu Asp Ser Val Val Ala Ser Asp Phe
1 5 10 15
Ile Ala Leu Gly Ile Thr Ser Glu Val Ala Glu Thr Leu His Gly Arg
20 25 30
Leu Ala Glu Ile Val Cys Asn Tyr Gly Ala Ala Thr Pro Gln Thr Trp
35 40 45
Ile Asn Ile Ala Asn His Ile Leu Ser Pro Asp Leu Pro Phe Ser Leu
50 55 60
His Gln Met Leu Phe Tyr Gly Cys Tyr Lys Asp Phe Gly Pro Ala Pro
65 70 75 80
Pro Ala Trp Ile Pro Asp Pro Glu Lys Val Lys Ser Thr Asn Leu Gly
85 90 95
Ala Leu Leu Glu Lys Arg Gly Lys Glu Phe Leu Gly Val Lys Tyr Lys
100 105 110
Asp Pro Ile Ser Ser Phe Ser His Phe Gln Glu Phe Ser Val Arg Asn
115 120 125
Pro Glu Val Tyr Trp Arg Thr Val Leu Met Asp Glu Met Lys Ile Ser
130 135 140
Phe Ser Lys Asp Pro Glu Cys Ile Leu Arg Arg Asp Asp Ile Asn Asn
145 150 155 160
Pro Gly Gly Ser Glu Trp Leu Pro Gly Gly Tyr Leu Asn Ser Ala Lys
165 170 175
Asn Cys Leu Asn Val Asn Ser Asn Lys Lys Leu Asn Asp Thr Met Ile
180 185 190
Val Trp Arg Asp Glu Gly Asn Asp Asp Leu Pro Leu Asn Lys Leu Thr
195 200 205
Leu Asp Gln Leu Arg Lys Arg Val Trp Leu Val Gly Tyr Ala Leu Glu
210 215 220
Glu Met Gly Leu Glu Lys Gly Cys Ala Ile Ala Ile Asp Met Pro Met
225 230 235 240
His Val Asp Ala Val Val Ile Tyr Leu Ala Ile Val Leu Ala Gly Tyr
245 250 255
Val Val Val Ser Ile Ala Asp Ser Phe Ser Ala Pro Glu Ile Ser Thr
260 265 270
Arg Leu Arg Leu Ser Lys Ala Lys Ala Ile Phe Thr Gln Asp His Ile
275 280 285
Ile Arg Gly Lys Lys Arg Ile Pro Leu Tyr Ser Arg Val Val Glu Ala
290 295 300
Lys Ser Pro Met Ala Ile Val Ile Pro Cys Ser Gly Ser Asn Ile Gly
305 310 315 320
Ala Glu Leu Arg Asp Gly Asp Ile Ser Trp Asp Tyr Phe Leu Glu Arg
325 330 335
Ala Lys Glu Phe Lys Asn Cys Glu Phe Thr Ala Arg Glu Gln Pro Val
340 345 350
Asp Ala Tyr Thr Asn Ile Leu Phe Ser Ser Gly Thr Thr Gly Glu Pro
355 360 365
Lys Ala Ile Pro Trp Thr Gln Ala Thr Pro Leu Lys Ala Ala Ala Asp
370 375 380
Gly Trp Ser His Leu Asp Ile Arg Lys Gly Asp Val Ile Val Trp Pro
385 390 395 400
Thr Asn Leu Gly Trp Met Met Gly Pro Trp Leu Val Tyr Ala Ser Leu
405 410 415
Leu Asn Gly Ala Ser Ile Ala Leu Tyr Asn Gly Ser Pro Leu Val Ser
420 425 430
Gly Phe Ala Lys Phe Val Gln Asp Ala Lys Val Thr Met Leu Gly Val
435 440 445
Val Pro Ser Ile Val Arg Ser Trp Lys Ser Thr Asn Cys Val Ser Gly
450 455 460
Tyr Asp Trp Ser Thr Ile Arg Cys Phe Ser Ser Ser Gly Glu Ala Ser
465 470 475 480
Asn Val Asp Glu Tyr Leu Trp Leu Met Gly Arg Ala Asn Tyr Lys Pro
485 490 495
Val Ile Glu Met Cys Gly Gly Thr Glu Ile Gly Gly Ala Phe Ser Ala
500 505 510
Gly Ser Phe Leu Gln Ala Gln Ser Leu Ser Ser Phe Ser Ser Gln Cys
515 520 525
Met Gly Cys Thr Leu Tyr Ile Leu Asp Lys Asn Gly Tyr Pro Met Pro
530 535 540
Lys Asn Lys Pro Gly Ile Gly Glu Leu Ala Leu Gly Pro Val Met Phe
545 550 555 560
Gly Ala Ser Lys Thr Leu Leu Asn Gly Asn His His Asp Val Tyr Phe
565 570 575
Lys Gly Met Pro Thr Leu Asn Gly Glu Val Leu Arg Arg His Gly Asp
580 585 590
Ile Phe Glu Leu Thr Ser Asn Gly Tyr Tyr His Ala His Gly Arg Ala
595 600 605
Asp Asp Thr Met Asn Ile Gly Gly Ile Lys Ile Ser Ser Ile Glu Ile
610 615 620
Glu Arg Val Cys Asn Glu Val Asp Asp Arg Val Phe Glu Thr Thr Ala
625 630 635 640
Ile Gly Val Pro Pro Leu Gly Gly Gly Pro Glu Gln Leu Val Ile Phe
645 650 655
Phe Val Leu Lys Asp Ser Asn Asp Thr Thr Ile Asp Leu Asn Gln Leu
660 665 670
Arg Leu Ser Phe Asn Leu Gly Leu Gln Lys Lys Leu Asn Pro Leu Phe
675 680 685
Lys Val Thr Arg Val Val Pro Leu Ser Ser Leu Pro Arg Thr Ala Thr
690 695 700
Asn Lys Ile Met Arg Arg Val Leu Arg Gln Gln Phe Ser His Phe Glu
705 710 715 720
<210> 48
<211> 2163
<212> DNA
<213> 大麻
<400> 48
atgggtaaga actacaagtc tttggactct gttgttgctt ctgacttcat cgctttgggt 60
atcacttctg aagttgctga aactttgcac ggtagattgg ctgaaatcgt ttgtaactac 120
ggtgctgcta ctccacaaac ttggatcaac atcgctaacc acatcttgtc tccagacttg 180
ccattctctt tgcaccaaat gttgttctac ggttgttaca aggacttcgg tccagctcca 240
ccagcttgga tcccagaccc agaaaaggtt aagtctacta acttgggtgc tttgttggaa 300
aagagaggta aggaattctt gggtgttaag tacaaggacc caatctcttc tttctctcac 360
ttccaagaat tctctgttag aaacccagaa gtttactgga gaactgtttt gatggacgaa 420
atgaagatct ctttctctaa ggacccagaa tgtatcttga gaagagacga catcaacaac 480
ccaggtggtt ctgaatggtt gccaggtggt tacttgaact ctgctaagaa ctgtttgaac 540
gttaactcta acaagaagtt gaacgacact atgatcgttt ggagagacga aggtaacgac 600
gacttgccat tgaacaagtt gactttggac caattgagaa agagagtttg gttggttggt 660
tacgctttgg aagaaatggg tttggaaaag ggttgtgcta tcgctatcga catgccaatg 720
cacgttgacg ctgttgttat ctacttggct atcgttttgg ctggttacgt tgttgtttct 780
atcgctgact ctttctctgc tccagaaatc tctactagat tgagattgtc taaggctaag 840
gctatcttca ctcaagacca catcatcaga ggtaagaaga gaatcccatt gtactctaga 900
gttgttgaag ctaagtctcc aatggctatc gttatcccat gttctggttc taacatcggt 960
gctgaattga gagacggtga catctcttgg gactacttct tggaaagagc taaggaattc 1020
aagaactgtg aattcactgc tagagaacaa ccagttgacg cttacactaa catcttgttc 1080
tcttctggta ctactggtga accaaaggct atcccatgga ctcaagctac tccattgaag 1140
gctgctgctg acggttggtc tcacttggac atcagaaagg gtgacgttat cgtttggcca 1200
actaacttgg gttggatgat gggtccatgg ttggtttacg cttctttgtt gaacggtgct 1260
tctatcgctt tgtacaacgg ttctccattg gtttctggtt tcgctaagtt cgttcaagac 1320
gctaaggtta ctatgttggg tgttgttcca tctatcgtta gatcttggaa gtctactaac 1380
tgtgtttctg gttacgactg gtctactatc agatgtttct cttcttctgg tgaagcttct 1440
aacgttgacg aatacttgtg gttgatgggt agagctaact acaagccagt tatcgaaatg 1500
tgtggtggta ctgaaatcgg tggtgctttc tctgctggtt ctttcttgca agctcaatct 1560
ttgtcttctt tctcttctca atgtatgggt tgtactttgt acatcttgga caagaacggt 1620
tacccaatgc caaagaacaa gccaggtatc ggtgaattgg ctttgggtcc agttatgttc 1680
ggtgcttcta agactttgtt gaacggtaac caccacgacg tttacttcaa gggtatgcca 1740
actttgaacg gtgaagtttt gagaagacac ggtgacatct tcgaattgac ttctaacggt 1800
tactaccacg ctcacggtag agctgacgac actatgaaca tcggtggtat caagatctct 1860
tctatcgaaa tcgaaagagt ttgtaacgaa gttgacgaca gagttttcga aactactgct 1920
atcggtgttc caccattggg tggtggtcca gaacaattgg ttatcttctt cgttttgaag 1980
gactctaacg acactactat cgacttgaac caattgagat tgtctttcaa cttgggtttg 2040
caaaagaagt tgaacccatt gttcaaggtt actagagttg ttccattgtc ttctttgcca 2100
agaactgcta ctaacaagat catgagaaga gttttgagac aacaattctc tcacttcgaa 2160
tag 2163
<210> 49
<211> 385
<212> PRT
<213> 大麻
<400> 49
Met Asn His Leu Arg Ala Glu Gly Pro Ala Ser Val Leu Ala Ile Gly
1 5 10 15
Thr Ala Asn Pro Glu Asn Ile Leu Leu Gln Asp Glu Phe Pro Asp Tyr
20 25 30
Tyr Phe Arg Val Thr Lys Ser Glu His Met Thr Gln Leu Lys Glu Lys
35 40 45
Phe Arg Lys Ile Cys Asp Lys Ser Met Ile Arg Lys Arg Asn Cys Phe
50 55 60
Leu Asn Glu Glu His Leu Lys Gln Asn Pro Arg Leu Val Glu His Glu
65 70 75 80
Met Gln Thr Leu Asp Ala Arg Gln Asp Met Leu Val Val Glu Val Pro
85 90 95
Lys Leu Gly Lys Asp Ala Cys Ala Lys Ala Ile Lys Glu Trp Gly Gln
100 105 110
Pro Lys Ser Lys Ile Thr His Leu Ile Phe Thr Ser Ala Ser Thr Thr
115 120 125
Asp Met Pro Gly Ala Asp Tyr His Cys Ala Lys Leu Leu Gly Leu Ser
130 135 140
Pro Ser Val Lys Arg Val Met Met Tyr Gln Leu Gly Cys Tyr Gly Gly
145 150 155 160
Gly Thr Val Leu Arg Ile Ala Lys Asp Ile Ala Glu Asn Asn Lys Gly
165 170 175
Ala Arg Val Leu Ala Val Cys Cys Asp Ile Met Ala Cys Leu Phe Arg
180 185 190
Gly Pro Ser Glu Ser Asp Leu Glu Leu Leu Val Gly Gln Ala Ile Phe
195 200 205
Gly Asp Gly Ala Ala Ala Val Ile Val Gly Ala Glu Pro Asp Glu Ser
210 215 220
Val Gly Glu Arg Pro Ile Phe Glu Leu Val Ser Thr Gly Gln Thr Ile
225 230 235 240
Leu Pro Asn Ser Glu Gly Thr Ile Gly Gly His Ile Arg Glu Ala Gly
245 250 255
Leu Ile Phe Asp Leu His Lys Asp Val Pro Met Leu Ile Ser Asn Asn
260 265 270
Ile Glu Lys Cys Leu Ile Glu Ala Phe Thr Pro Ile Gly Ile Ser Asp
275 280 285
Trp Asn Ser Ile Phe Trp Ile Thr His Pro Gly Gly Lys Ala Ile Leu
290 295 300
Asp Lys Val Glu Glu Lys Leu His Leu Lys Ser Asp Lys Phe Val Asp
305 310 315 320
Ser Arg His Val Leu Ser Glu His Gly Asn Met Ser Ser Ser Thr Val
325 330 335
Leu Phe Val Met Asp Glu Leu Arg Lys Arg Ser Leu Glu Glu Gly Lys
340 345 350
Ser Thr Thr Gly Asp Gly Phe Glu Trp Gly Val Leu Phe Gly Phe Gly
355 360 365
Pro Gly Leu Thr Val Glu Arg Val Val Val Arg Ser Val Pro Ile Lys
370 375 380
Tyr
385
<210> 50
<211> 1158
<212> DNA
<213> 大麻
<400> 50
atgaaccact tgagagctga aggtccagct tctgttttgg ctatcggtac tgctaaccca 60
gaaaacatct tgttgcaaga cgaattccca gactactact tcagagttac taagtctgaa 120
cacatgactc aattgaagga aaagttcaga aagatctgtg acaagtctat gatcagaaag 180
agaaactgtt tcttgaacga agaacacttg aagcaaaacc caagattggt tgaacacgaa 240
atgcaaactt tggacgctag acaagacatg ttggttgttg aagttccaaa gttgggtaag 300
gacgcttgtg ctaaggctat caaggaatgg ggtcaaccaa agtctaagat cactcacttg 360
atcttcactt ctgcttctac tactgacatg ccaggtgctg actaccactg tgctaagttg 420
ttgggtttgt ctccatctgt taagagagtt atgatgtacc aattgggttg ttacggtggt 480
ggtactgttt tgagaatcgc taaggacatc gctgaaaaca acaagggtgc tagagttttg 540
gctgtttgtt gtgacatcat ggcttgtttg ttcagaggtc catctgaatc tgacttggaa 600
ttgttggttg gtcaagctat cttcggtgac ggtgctgctg ctgttatcgt tggtgctgaa 660
ccagacgaat ctgttggtga aagaccaatc ttcgaattgg tttctactgg tcaaactatc 720
ttgccaaact ctgaaggtac tatcggtggt cacatcagag aagctggttt gatcttcgac 780
ttgcacaagg acgttccaat gttgatctct aacaacatcg aaaagtgttt gatcgaagct 840
ttcactccaa tcggtatctc tgactggaac tctatcttct ggatcactca cccaggtggt 900
aaggctatct tggacaaggt tgaagaaaag ttgcacttga agtctgacaa gttcgttgac 960
tctagacacg ttttgtctga acacggtaac atgtcttctt ctactgtttt gttcgttatg 1020
gacgaattga gaaagagatc tttggaagaa ggtaagtcta ctactggtga cggtttcgaa 1080
tggggtgttt tgttcggttt cggtccaggt ttgactgttg aaagagttgt tgttagatct 1140
gttccaatca agtactag 1158
<210> 51
<211> 101
<212> PRT
<213> 大麻
<400> 51
Met Ala Val Lys His Leu Ile Val Leu Lys Phe Lys Asp Glu Ile Thr
1 5 10 15
Glu Ala Gln Lys Glu Glu Phe Phe Lys Thr Tyr Val Asn Leu Val Asn
20 25 30
Ile Ile Pro Ala Met Lys Asp Val Tyr Trp Gly Lys Asp Val Thr Gln
35 40 45
Lys Asn Lys Glu Glu Gly Tyr Thr His Ile Val Glu Val Thr Phe Glu
50 55 60
Ser Val Glu Thr Ile Gln Asp Tyr Ile Ile His Pro Ala His Val Gly
65 70 75 80
Phe Gly Asp Val Tyr Arg Ser Phe Trp Glu Lys Leu Leu Ile Phe Asp
85 90 95
Tyr Thr Pro Arg Lys
100
<210> 52
<211> 306
<212> DNA
<213> 大麻
<400> 52
atggctgtta agcacttgat cgttttgaag ttcaaggacg aaatcactga agctcaaaag 60
gaagaattct tcaagactta cgttaacttg gttaacatca tcccagctat gaaggacgtt 120
tactggggta aggacgttac tcaaaagaac aaggaagaag gttacactca catcgttgaa 180
gttactttcg aatctgttga aactatccaa gactacatca tccacccagc tcacgttggt 240
ttcggtgacg tttacagatc tttctgggaa aagttgttga tcttcgacta cactccaaga 300
aagtag 306
<210> 53
<211> 398
<212> PRT
<213> 大麻
<400> 53
Met Gly Leu Ser Leu Val Cys Thr Phe Ser Phe Gln Thr Asn Tyr His
1 5 10 15
Thr Leu Leu Asn Pro His Asn Lys Asn Pro Lys Asn Ser Leu Leu Ser
20 25 30
Tyr Gln His Pro Lys Thr Pro Ile Ile Lys Ser Ser Tyr Asp Asn Phe
35 40 45
Pro Ser Lys Tyr Cys Leu Thr Lys Asn Phe His Leu Leu Gly Leu Asn
50 55 60
Ser His Asn Arg Ile Ser Ser Gln Ser Arg Ser Ile Arg Ala Gly Ser
65 70 75 80
Asp Gln Ile Glu Gly Ser Pro His His Glu Ser Asp Asn Ser Ile Ala
85 90 95
Thr Lys Ile Leu Asn Phe Gly His Thr Cys Trp Lys Leu Gln Arg Pro
100 105 110
Tyr Val Val Lys Gly Met Ile Ser Ile Ala Cys Gly Leu Phe Gly Arg
115 120 125
Glu Leu Phe Asn Asn Arg His Leu Phe Ser Trp Gly Leu Met Trp Lys
130 135 140
Ala Phe Phe Ala Leu Val Pro Ile Leu Ser Phe Asn Phe Phe Ala Ala
145 150 155 160
Ile Met Asn Gln Ile Tyr Asp Val Asp Ile Asp Arg Ile Asn Lys Pro
165 170 175
Asp Leu Pro Leu Val Ser Gly Glu Met Ser Ile Glu Thr Ala Trp Ile
180 185 190
Leu Ser Ile Ile Val Ala Leu Thr Gly Leu Ile Val Thr Ile Lys Leu
195 200 205
Lys Ser Ala Pro Leu Phe Val Phe Ile Tyr Ile Phe Gly Ile Phe Ala
210 215 220
Gly Phe Ala Tyr Ser Val Pro Pro Ile Arg Trp Lys Gln Tyr Pro Phe
225 230 235 240
Thr Asn Phe Leu Ile Thr Ile Ser Ser His Val Gly Leu Ala Phe Thr
245 250 255
Ser Tyr Ser Ala Thr Thr Ser Ala Leu Gly Leu Pro Phe Val Trp Arg
260 265 270
Pro Ala Phe Ser Phe Ile Ile Ala Phe Met Thr Val Met Gly Met Thr
275 280 285
Ile Ala Phe Ala Lys Asp Ile Ser Asp Ile Glu Gly Asp Ala Lys Tyr
290 295 300
Gly Val Ser Thr Val Ala Thr Lys Leu Gly Ala Arg Asn Met Thr Phe
305 310 315 320
Val Val Ser Gly Val Leu Leu Leu Asn Tyr Leu Val Ser Ile Ser Ile
325 330 335
Gly Ile Ile Trp Pro Gln Val Phe Lys Ser Asn Ile Met Ile Leu Ser
340 345 350
His Ala Ile Leu Ala Phe Cys Leu Ile Phe Gln Thr Arg Glu Leu Ala
355 360 365
Leu Ala Asn Tyr Ala Ser Ala Pro Ser Arg Gln Phe Phe Glu Phe Ile
370 375 380
Trp Leu Leu Tyr Tyr Ala Glu Tyr Phe Val Tyr Val Phe Ile
385 390 395
<210> 54
<211> 1197
<212> DNA
<213> 大麻
<400> 54
atgggtttgt ctttggtttg tactttctct ttccaaacta actaccacac tttgttgaac 60
ccacacaaca agaacccaaa gaactctttg ttgtcttacc aacacccaaa gactccaatc 120
atcaagtctt cttacgacaa cttcccatct aagtactgtt tgactaagaa cttccacttg 180
ttgggtttga actctcacaa cagaatctct tctcaatcta gatctatcag agctggttct 240
gaccaaatcg aaggttctcc acaccacgaa tctgacaact ctatcgctac taagatcttg 300
aacttcggtc acacttgttg gaagttgcaa agaccatacg ttgttaaggg tatgatctct 360
atcgcttgtg gtttgttcgg tagagaattg ttcaacaaca gacacttgtt ctcttggggt 420
ttgatgtgga aggctttctt cgctttggtt ccaatcttgt ctttcaactt cttcgctgct 480
atcatgaacc aaatctacga cgttgacatc gacagaatca acaagccaga cttgccattg 540
gtttctggtg aaatgtctat cgaaactgct tggatcttgt ctatcatcgt tgctttgact 600
ggtttgatcg ttactatcaa gttgaagtct gctccattgt tcgttttcat ctacatcttc 660
ggtatcttcg ctggtttcgc ttactctgtt ccaccaatca gatggaagca atacccattc 720
actaacttct tgatcactat ctcttctcac gttggtttgg ctttcacttc ttactctgct 780
actacttctg ctttgggttt gccattcgtt tggagaccag ctttctcttt catcatcgct 840
ttcatgactg ttatgggtat gactatcgct ttcgctaagg acatctctga catcgaaggt 900
gacgctaagt acggtgtttc tactgttgct actaagttgg gtgctagaaa catgactttc 960
gttgtttctg gtgttttgtt gttgaactac ttggtttcta tctctatcgg tatcatctgg 1020
ccacaagttt tcaagtctaa catcatgatc ttgtctcacg ctatcttggc tttctgtttg 1080
atcttccaaa ctagagaatt ggctttggct aactacgctt ctgctccatc tagacaattc 1140
ttcgaattca tctggttgtt gtactacgct gaatacttcg tttacgtttt catctag 1197
<210> 55
<211> 545
<212> PRT
<213> 大麻
<400> 55
Met Asn Cys Ser Ala Phe Ser Phe Trp Phe Val Cys Lys Ile Ile Phe
1 5 10 15
Phe Phe Leu Ser Phe His Ile Gln Ile Ser Ile Ala Asn Pro Arg Glu
20 25 30
Asn Phe Leu Lys Cys Phe Ser Lys His Ile Pro Asn Asn Val Ala Asn
35 40 45
Pro Lys Leu Val Tyr Thr Gln His Asp Gln Leu Tyr Met Ser Ile Leu
50 55 60
Asn Ser Thr Ile Gln Asn Leu Arg Phe Ile Ser Asp Thr Thr Pro Lys
65 70 75 80
Pro Leu Val Ile Val Thr Pro Ser Asn Asn Ser His Ile Gln Ala Thr
85 90 95
Ile Leu Cys Ser Lys Lys Val Gly Leu Gln Ile Arg Thr Arg Ser Gly
100 105 110
Gly His Asp Ala Glu Gly Met Ser Tyr Ile Ser Gln Val Pro Phe Val
115 120 125
Val Val Asp Leu Arg Asn Met His Ser Ile Lys Ile Asp Val His Ser
130 135 140
Gln Thr Ala Trp Val Glu Ala Gly Ala Thr Leu Gly Glu Val Tyr Tyr
145 150 155 160
Trp Ile Asn Glu Lys Asn Glu Asn Leu Ser Phe Pro Gly Gly Tyr Cys
165 170 175
Pro Thr Val Gly Val Gly Gly His Phe Ser Gly Gly Gly Tyr Gly Ala
180 185 190
Leu Met Arg Asn Tyr Gly Leu Ala Ala Asp Asn Ile Ile Asp Ala His
195 200 205
Leu Val Asn Val Asp Gly Lys Val Leu Asp Arg Lys Ser Met Gly Glu
210 215 220
Asp Leu Phe Trp Ala Ile Arg Gly Gly Gly Gly Glu Asn Phe Gly Ile
225 230 235 240
Ile Ala Ala Trp Lys Ile Lys Leu Val Ala Val Pro Ser Lys Ser Thr
245 250 255
Ile Phe Ser Val Lys Lys Asn Met Glu Ile His Gly Leu Val Lys Leu
260 265 270
Phe Asn Lys Trp Gln Asn Ile Ala Tyr Lys Tyr Asp Lys Asp Leu Val
275 280 285
Leu Met Thr His Phe Ile Thr Lys Asn Ile Thr Asp Asn His Gly Lys
290 295 300
Asn Lys Thr Thr Val His Gly Tyr Phe Ser Ser Ile Phe His Gly Gly
305 310 315 320
Val Asp Ser Leu Val Asp Leu Met Asn Lys Ser Phe Pro Glu Leu Gly
325 330 335
Ile Lys Lys Thr Asp Cys Lys Glu Phe Ser Trp Ile Asp Thr Thr Ile
340 345 350
Phe Tyr Ser Gly Val Val Asn Phe Asn Thr Ala Asn Phe Lys Lys Glu
355 360 365
Ile Leu Leu Asp Arg Ser Ala Gly Lys Lys Thr Ala Phe Ser Ile Lys
370 375 380
Leu Asp Tyr Val Lys Lys Pro Ile Pro Glu Thr Ala Met Val Lys Ile
385 390 395 400
Leu Glu Lys Leu Tyr Glu Glu Asp Val Gly Ala Gly Met Tyr Val Leu
405 410 415
Tyr Pro Tyr Gly Gly Ile Met Glu Glu Ile Ser Glu Ser Ala Ile Pro
420 425 430
Phe Pro His Arg Ala Gly Ile Met Tyr Glu Leu Trp Tyr Thr Ala Ser
435 440 445
Trp Glu Lys Gln Glu Asp Asn Glu Lys His Ile Asn Trp Val Arg Ser
450 455 460
Val Tyr Asn Phe Thr Thr Pro Tyr Val Ser Gln Asn Pro Arg Leu Ala
465 470 475 480
Tyr Leu Asn Tyr Arg Asp Leu Asp Leu Gly Lys Thr Asn His Ala Ser
485 490 495
Pro Asn Asn Tyr Thr Gln Ala Arg Ile Trp Gly Glu Lys Tyr Phe Gly
500 505 510
Lys Asn Phe Asn Arg Leu Val Lys Val Lys Thr Lys Val Asp Pro Asn
515 520 525
Asn Phe Phe Arg Asn Glu Gln Ser Ile Pro Pro Leu Pro Pro His His
530 535 540
His
545
<210> 56
<211> 1638
<212> DNA
<213> 大麻
<400> 56
atgaactgtt ctgctttctc tttctggttc gtttgtaaga tcatcttctt cttcttgtct 60
ttccacatcc aaatctctat cgctaaccca agagaaaact tcttgaagtg tttctctaag 120
cacatcccaa acaacgttgc taacccaaag ttggtttaca ctcaacacga ccaattgtac 180
atgtctatct tgaactctac tatccaaaac ttgagattca tctctgacac tactccaaag 240
ccattggtta tcgttactcc atctaacaac tctcacatcc aagctactat cttgtgttct 300
aagaaggttg gtttgcaaat cagaactaga tctggtggtc acgacgctga aggtatgtct 360
tacatctctc aagttccatt cgttgttgtt gacttgagaa acatgcactc tatcaagatc 420
gacgttcact ctcaaactgc ttgggttgaa gctggtgcta ctttgggtga agtttactac 480
tggatcaacg aaaagaacga aaacttgtct ttcccaggtg gttactgtcc aactgttggt 540
gttggtggtc acttctctgg tggtggttac ggtgctttga tgagaaacta cggtttggct 600
gctgacaaca tcatcgacgc tcacttggtt aacgttgacg gtaaggtttt ggacagaaag 660
tctatgggtg aagacttgtt ctgggctatc agaggtggtg gtggtgaaaa cttcggtatc 720
atcgctgctt ggaagatcaa gttggttgct gttccatcta agtctactat cttctctgtt 780
aagaagaaca tggaaatcca cggtttggtt aagttgttca acaagtggca aaacatcgct 840
tacaagtacg acaaggactt ggttttgatg actcacttca tcactaagaa catcactgac 900
aaccacggta agaacaagac tactgttcac ggttacttct cttctatctt ccacggtggt 960
gttgactctt tggttgactt gatgaacaag tctttcccag aattgggtat caagaagact 1020
gactgtaagg aattctcttg gatcgacact actatcttct actctggtgt tgttaacttc 1080
aacactgcta acttcaagaa ggaaatcttg ttggacagat ctgctggtaa gaagactgct 1140
ttctctatca agttggacta cgttaagaag ccaatcccag aaactgctat ggttaagatc 1200
ttggaaaagt tgtacgaaga agacgttggt gctggtatgt acgttttgta cccatacggt 1260
ggtatcatgg aagaaatctc tgaatctgct atcccattcc cacacagagc tggtatcatg 1320
tacgaattgt ggtacactgc ttcttgggaa aagcaagaag acaacgaaaa gcacatcaac 1380
tgggttagat ctgtttacaa cttcactact ccatacgttt ctcaaaaccc aagattggct 1440
tacttgaact acagagactt ggacttgggt aagactaacc acgcttctcc aaacaactac 1500
actcaagcta gaatctgggg tgaaaagtac ttcggtaaga acttcaacag attggttaag 1560
gttaagacta aggttgaccc aaacaacttc ttcagaaacg aacaatctat cccaccattg 1620
ccaccacacc accactag 1638
<210> 57
<211> 544
<212> PRT
<213> 大麻
<400> 57
Met Lys Cys Ser Thr Phe Ser Phe Trp Phe Val Cys Lys Ile Ile Phe
1 5 10 15
Phe Phe Phe Ser Phe Asn Ile Gln Thr Ser Ile Ala Asn Pro Arg Glu
20 25 30
Asn Phe Leu Lys Cys Phe Ser Gln Tyr Ile Pro Asn Asn Ala Thr Asn
35 40 45
Leu Lys Leu Val Tyr Thr Gln Asn Asn Pro Leu Tyr Met Ser Val Leu
50 55 60
Asn Ser Thr Ile His Asn Leu Arg Phe Thr Ser Asp Thr Thr Pro Lys
65 70 75 80
Pro Leu Val Ile Val Thr Pro Ser His Val Ser His Ile Gln Gly Thr
85 90 95
Ile Leu Cys Ser Lys Lys Val Gly Leu Gln Ile Arg Thr Arg Ser Gly
100 105 110
Gly His Asp Ser Glu Gly Met Ser Tyr Ile Ser Gln Val Pro Phe Val
115 120 125
Ile Val Asp Leu Arg Asn Met Arg Ser Ile Lys Ile Asp Val His Ser
130 135 140
Gln Thr Ala Trp Val Glu Ala Gly Ala Thr Leu Gly Glu Val Tyr Tyr
145 150 155 160
Trp Val Asn Glu Lys Asn Glu Asn Leu Ser Leu Ala Ala Gly Tyr Cys
165 170 175
Pro Thr Val Cys Ala Gly Gly His Phe Gly Gly Gly Gly Tyr Gly Pro
180 185 190
Leu Met Arg Asn Tyr Gly Leu Ala Ala Asp Asn Ile Ile Asp Ala His
195 200 205
Leu Val Asn Val His Gly Lys Val Leu Asp Arg Lys Ser Met Gly Glu
210 215 220
Asp Leu Phe Trp Ala Leu Arg Gly Gly Gly Ala Glu Ser Phe Gly Ile
225 230 235 240
Ile Val Ala Trp Lys Ile Arg Leu Val Ala Val Pro Lys Ser Thr Met
245 250 255
Phe Ser Val Lys Lys Ile Met Glu Ile His Glu Leu Val Lys Leu Val
260 265 270
Asn Lys Trp Gln Asn Ile Ala Tyr Lys Tyr Asp Lys Asp Leu Leu Leu
275 280 285
Met Thr His Phe Ile Thr Arg Asn Ile Thr Asp Asn Gln Gly Lys Asn
290 295 300
Lys Thr Ala Ile His Thr Tyr Phe Ser Ser Val Phe Leu Gly Gly Val
305 310 315 320
Asp Ser Leu Val Asp Leu Met Asn Lys Ser Phe Pro Glu Leu Gly Ile
325 330 335
Lys Lys Thr Asp Cys Arg Gln Leu Ser Trp Ile Asp Thr Ile Ile Phe
340 345 350
Tyr Ser Gly Val Val Asn Tyr Asp Thr Asp Asn Phe Asn Lys Glu Ile
355 360 365
Leu Leu Asp Arg Ser Ala Gly Gln Asn Gly Ala Phe Lys Ile Lys Leu
370 375 380
Asp Tyr Val Lys Lys Pro Ile Pro Glu Ser Val Phe Val Gln Ile Leu
385 390 395 400
Glu Lys Leu Tyr Glu Glu Asp Ile Gly Ala Gly Met Tyr Ala Leu Tyr
405 410 415
Pro Tyr Gly Gly Ile Met Asp Glu Ile Ser Glu Ser Ala Ile Pro Phe
420 425 430
Pro His Arg Ala Gly Ile Leu Tyr Glu Leu Trp Tyr Ile Cys Ser Trp
435 440 445
Glu Lys Gln Glu Asp Asn Glu Lys His Leu Asn Trp Ile Arg Asn Ile
450 455 460
Tyr Asn Phe Met Thr Pro Tyr Val Ser Lys Asn Pro Arg Leu Ala Tyr
465 470 475 480
Leu Asn Tyr Arg Asp Leu Asp Ile Gly Ile Asn Asp Pro Lys Asn Pro
485 490 495
Asn Asn Tyr Thr Gln Ala Arg Ile Trp Gly Glu Lys Tyr Phe Gly Lys
500 505 510
Asn Phe Asp Arg Leu Val Lys Val Lys Thr Leu Val Asp Pro Asn Asn
515 520 525
Phe Phe Arg Asn Glu Gln Ser Ile Pro Pro Leu Pro Arg His Arg His
530 535 540
<210> 58
<211> 1635
<212> DNA
<213> 大麻
<400> 58
atgaagtgtt ctactttctc tttctggttc gtttgtaaga tcatcttctt cttcttctct 60
ttcaacatcc aaacttctat cgctaaccca agagaaaact tcttgaagtg tttctctcaa 120
tacatcccaa acaacgctac taacttgaag ttggtttaca ctcaaaacaa cccattgtac 180
atgtctgttt tgaactctac tatccacaac ttgagattca cttctgacac tactccaaag 240
ccattggtta tcgttactcc atctcacgtt tctcacatcc aaggtactat cttgtgttct 300
aagaaggttg gtttgcaaat cagaactaga tctggtggtc acgactctga aggtatgtct 360
tacatctctc aagttccatt cgttatcgtt gacttgagaa acatgagatc tatcaagatc 420
gacgttcact ctcaaactgc ttgggttgaa gctggtgcta ctttgggtga agtttactac 480
tgggttaacg aaaagaacga aaacttgtct ttggctgctg gttactgtcc aactgtttgt 540
gctggtggtc acttcggtgg tggtggttac ggtccattga tgagaaacta cggtttggct 600
gctgacaaca tcatcgacgc tcacttggtt aacgttcacg gtaaggtttt ggacagaaag 660
tctatgggtg aagacttgtt ctgggctttg agaggtggtg gtgctgaatc tttcggtatc 720
atcgttgctt ggaagatcag attggttgct gttccaaagt ctactatgtt ctctgttaag 780
aagatcatgg aaatccacga attggttaag ttggttaaca agtggcaaaa catcgcttac 840
aagtacgaca aggacttgtt gttgatgact cacttcatca ctagaaacat cactgacaac 900
caaggtaaga acaagactgc tatccacact tacttctctt ctgttttctt gggtggtgtt 960
gactctttgg ttgacttgat gaacaagtct ttcccagaat tgggtatcaa gaagactgac 1020
tgtagacaat tgtcttggat cgacactatc atcttctact ctggtgttgt taactacgac 1080
actgacaact tcaacaagga aatcttgttg gacagatctg ctggtcaaaa cggtgctttc 1140
aagatcaagt tggactacgt taagaagcca atcccagaat ctgttttcgt tcaaatcttg 1200
gaaaagttgt acgaagaaga catcggtgct ggtatgtacg ctttgtaccc atacggtggt 1260
atcatggacg aaatctctga atctgctatc ccattcccac acagagctgg tatcttgtac 1320
gaattgtggt acatctgttc ttgggaaaag caagaagaca acgaaaagca cttgaactgg 1380
atcagaaaca tctacaactt catgactcca tacgtttcta agaacccaag attggcttac 1440
ttgaactaca gagacttgga catcggtatc aacgacccaa agaacccaaa caactacact 1500
caagctagaa tctggggtga aaagtacttc ggtaagaact tcgacagatt ggttaaggtt 1560
aagactttgg ttgacccaaa caacttcttc agaaacgaac aatctatccc accattgcca 1620
agacacagac actag 1635
<210> 59
<211> 545
<212> PRT
<213> 大麻
<400> 59
Met Asn Cys Ser Thr Phe Ser Phe Trp Phe Val Cys Lys Ile Ile Phe
1 5 10 15
Phe Phe Leu Ser Phe Asn Ile Gln Ile Ser Ile Ala Asn Pro Gln Glu
20 25 30
Asn Phe Leu Lys Cys Phe Ser Glu Tyr Ile Pro Asn Asn Pro Ala Asn
35 40 45
Pro Lys Phe Ile Tyr Thr Gln His Asp Gln Leu Tyr Met Ser Val Leu
50 55 60
Asn Ser Thr Ile Gln Asn Leu Arg Phe Thr Ser Asp Thr Thr Pro Lys
65 70 75 80
Pro Leu Val Ile Val Thr Pro Ser Asn Val Ser His Ile Gln Ala Ser
85 90 95
Ile Leu Cys Ser Lys Lys Val Gly Leu Gln Ile Arg Thr Arg Ser Gly
100 105 110
Gly His Asp Ala Glu Gly Leu Ser Tyr Ile Ser Gln Val Pro Phe Ala
115 120 125
Ile Val Asp Leu Arg Asn Met His Thr Val Lys Val Asp Ile His Ser
130 135 140
Gln Thr Ala Trp Val Glu Ala Gly Ala Thr Leu Gly Glu Val Tyr Tyr
145 150 155 160
Trp Ile Asn Glu Met Asn Glu Asn Phe Ser Phe Pro Gly Gly Tyr Cys
165 170 175
Pro Thr Val Gly Val Gly Gly His Phe Ser Gly Gly Gly Tyr Gly Ala
180 185 190
Leu Met Arg Asn Tyr Gly Leu Ala Ala Asp Asn Ile Ile Asp Ala His
195 200 205
Leu Val Asn Val Asp Gly Lys Val Leu Asp Arg Lys Ser Met Gly Glu
210 215 220
Asp Leu Phe Trp Ala Ile Arg Gly Gly Gly Gly Glu Asn Phe Gly Ile
225 230 235 240
Ile Ala Ala Cys Lys Ile Lys Leu Val Val Val Pro Ser Lys Ala Thr
245 250 255
Ile Phe Ser Val Lys Lys Asn Met Glu Ile His Gly Leu Val Lys Leu
260 265 270
Phe Asn Lys Trp Gln Asn Ile Ala Tyr Lys Tyr Asp Lys Asp Leu Met
275 280 285
Leu Thr Thr His Phe Arg Thr Arg Asn Ile Thr Asp Asn His Gly Lys
290 295 300
Asn Lys Thr Thr Val His Gly Tyr Phe Ser Ser Ile Phe Leu Gly Gly
305 310 315 320
Val Asp Ser Leu Val Asp Leu Met Asn Lys Ser Phe Pro Glu Leu Gly
325 330 335
Ile Lys Lys Thr Asp Cys Lys Glu Leu Ser Trp Ile Asp Thr Thr Ile
340 345 350
Phe Tyr Ser Gly Val Val Asn Tyr Asn Thr Ala Asn Phe Lys Lys Glu
355 360 365
Ile Leu Leu Asp Arg Ser Ala Gly Lys Lys Thr Ala Phe Ser Ile Lys
370 375 380
Leu Asp Tyr Val Lys Lys Leu Ile Pro Glu Thr Ala Met Val Lys Ile
385 390 395 400
Leu Glu Lys Leu Tyr Glu Glu Glu Val Gly Val Gly Met Tyr Val Leu
405 410 415
Tyr Pro Tyr Gly Gly Ile Met Asp Glu Ile Ser Glu Ser Ala Ile Pro
420 425 430
Phe Pro His Arg Ala Gly Ile Met Tyr Glu Leu Trp Tyr Thr Ala Thr
435 440 445
Trp Glu Lys Gln Glu Asp Asn Glu Lys His Ile Asn Trp Val Arg Ser
450 455 460
Val Tyr Asn Phe Thr Thr Pro Tyr Val Ser Gln Asn Pro Arg Leu Ala
465 470 475 480
Tyr Leu Asn Tyr Arg Asp Leu Asp Leu Gly Lys Thr Asn Pro Glu Ser
485 490 495
Pro Asn Asn Tyr Thr Gln Ala Arg Ile Trp Gly Glu Lys Tyr Phe Gly
500 505 510
Lys Asn Phe Asn Arg Leu Val Lys Val Lys Thr Lys Ala Asp Pro Asn
515 520 525
Asn Phe Phe Arg Asn Glu Gln Ser Ile Pro Pro Leu Pro Pro Arg His
530 535 540
His
545
<210> 60
<211> 1638
<212> DNA
<213> 大麻
<400> 60
atgaactgtt ctactttctc tttctggttc gtttgtaaga tcatcttctt cttcttgtct 60
ttcaacatcc aaatctctat cgctaaccca caagaaaact tcttgaagtg tttctctgaa 120
tacatcccaa acaacccagc taacccaaag ttcatctaca ctcaacacga ccaattgtac 180
atgtctgttt tgaactctac tatccaaaac ttgagattca cttctgacac tactccaaag 240
ccattggtta tcgttactcc atctaacgtt tctcacatcc aagcttctat cttgtgttct 300
aagaaggttg gtttgcaaat cagaactaga tctggtggtc acgacgctga aggtttgtct 360
tacatctctc aagttccatt cgctatcgtt gacttgagaa acatgcacac tgttaaggtt 420
gacatccact ctcaaactgc ttgggttgaa gctggtgcta ctttgggtga agtttactac 480
tggatcaacg aaatgaacga aaacttctct ttcccaggtg gttactgtcc aactgttggt 540
gttggtggtc acttctctgg tggtggttac ggtgctttga tgagaaacta cggtttggct 600
gctgacaaca tcatcgacgc tcacttggtt aacgttgacg gtaaggtttt ggacagaaag 660
tctatgggtg aagacttgtt ctgggctatc agaggtggtg gtggtgaaaa cttcggtatc 720
atcgctgctt gtaagatcaa gttggttgtt gttccatcta aggctactat cttctctgtt 780
aagaagaaca tggaaatcca cggtttggtt aagttgttca acaagtggca aaacatcgct 840
tacaagtacg acaaggactt gatgttgact actcacttca gaactagaaa catcactgac 900
aaccacggta agaacaagac tactgttcac ggttacttct cttctatctt cttgggtggt 960
gttgactctt tggttgactt gatgaacaag tctttcccag aattgggtat caagaagact 1020
gactgtaagg aattgtcttg gatcgacact actatcttct actctggtgt tgttaactac 1080
aacactgcta acttcaagaa ggaaatcttg ttggacagat ctgctggtaa gaagactgct 1140
ttctctatca agttggacta cgttaagaag ttgatcccag aaactgctat ggttaagatc 1200
ttggaaaagt tgtacgaaga agaagttggt gttggtatgt acgttttgta cccatacggt 1260
ggtatcatgg acgaaatctc tgaatctgct atcccattcc cacacagagc tggtatcatg 1320
tacgaattgt ggtacactgc tacttgggaa aagcaagaag acaacgaaaa gcacatcaac 1380
tgggttagat ctgtttacaa cttcactact ccatacgttt ctcaaaaccc aagattggct 1440
tacttgaact acagagactt ggacttgggt aagactaacc cagaatctcc aaacaactac 1500
actcaagcta gaatctgggg tgaaaagtac ttcggtaaga acttcaacag attggttaag 1560
gttaagacta aggctgaccc aaacaacttc ttcagaaacg aacaatctat cccaccattg 1620
ccaccaagac accactag 1638
<210> 61
<211> 26
<212> DNA
<213> 人工序列
<220>
<223> 人工
<400> 61
acctgcacut tgtaattaaa acttag 26
<210> 62
<211> 26
<212> DNA
<213> 人工序列
<220>
<223> 人工
<400> 62
atgacagaut tgttttatat ttgttg 26
<210> 63
<211> 37
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 63
agtgcaggua aaacaatggc tgttaagcac ttgatcg 37
<210> 64
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 64
cgtgcgauct ttcttggagt gtagtcgaag 30
<210> 65
<211> 38
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 65
atctgtcaua aaacaatgaa ccacttgaga gctgaagg 38
<210> 66
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 66
cacgcgaugt acttgattgg aacagatcta ac 32
<210> 67
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 67
acctgcacut ttgtttgttt atgtgtgttt attc 34
<210> 68
<211> 26
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 68
atgacagaut tgtaattaaa acttag 26
<210> 69
<211> 42
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 69
agtgcaggua aaacaatggg tttgtctttg gtttgtactt tc 42
<210> 70
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 70
cgtgcgauga tgaaaacgta aacgaagtat tc 32
<210> 71
<211> 40
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 71
atctgtcaua aaacaatgtt cgacttcaac aagtacatgg 40
<210> 72
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 72
cacgcgauct agttttgtct gaaagcaacg tag 33
<210> 73
<211> 25
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 73
cgtgcgaugg aagtaccttc aaaga 25
<210> 74
<211> 26
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 74
atgacagaut tgttttatat ttgttg 26
<210> 75
<211> 40
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 75
atctgtcaua aaacaatggg taagaactac aagtctttgg 40
<210> 76
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 76
cacgcgautt cgaagtgaga gaattgttgt ctc 33
<210> 77
<211> 26
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 77
acctgcacut tgtaattaaa acttag 26
<210> 78
<211> 25
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 78
cacgcgaugc acacaccata gcttc 25
<210> 79
<211> 42
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 79
agtgcaggua aaacaatgaa ctgttctgct ttctctttct gg 42
<210> 80
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 80
cgtgcgaugt ggtggtgtgg tggcaatgg 29
<210> 81
<211> 42
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 81
agtgcaggua aaacaatgaa gtgttctact ttctctttct gg 42
<210> 82
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 82
cgtgcgaugt gtctgtgtct tggcaatgg 29
<210> 83
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 83
agtgcaggua aaacaatgaa ctgttctact ttctctttc 39
<210> 84
<211> 29
<212> DNA
<213> Artificial
<220>
<223> 人工
<400> 84
cgtgcgaugt ggtgtcttgg tggcaatgg 29
<210> 85
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 85
ggatccatgg ctgttaagca cttgatcg 28
<210> 86
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 86
aagcttctac tttcttggag tgtagtcgaa g 31
<210> 87
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 87
cgccggcgat gaaccacttg agagctgaag g 31
<210> 88
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 88
cttaagctag tacttgattg gaacagatct aac 33
<210> 89
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 89
ggatccatgg gtttgtcttt ggtttgtact ttc 33
<210> 90
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 90
aagcttctag atgaaaacgt aaacgaagta ttc 33
<210> 91
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 91
cgccggcgat gttcgacttc aacaagtaca tgg 33
<210> 92
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 92
cttaagctac tagttttgtc tgaaagcaac gtag 34
<210> 93
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 93
ggatccatgg gtaagaacta caagtctttg g 31
<210> 94
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 94
aagcttctat tcgaagtgag agaattgttg tctc 34
<210> 95
<211> 35
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 95
cgccggcgat gaactgttct gctttctctt tctgg 35
<210> 96
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 96
cttaagctag tggtggtgtg gtggcaatgg 30
<210> 97
<211> 35
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 97
cgccggcgat gaagtgttct actttctctt tctgg 35
<210> 98
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 98
cttaagctag tgtctgtgtc ttggcaatgg 30
<210> 99
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 99
cgccggcgat gaactgttct actttctctt tc 32
<210> 100
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 100
cttaagctag tggtgtcttg gtggcaatgg 30
<210> 101
<211> 477
<212> PRT
<213> 毛果杨(P. trichocarpa)
<400> 101
Met Glu Asp Thr Ile Val Leu Tyr Pro Ser Pro Gly Arg Gly His Leu
1 5 10 15
Phe Ser Met Val Glu Leu Gly Lys Gln Ile Leu Glu His His Pro Ser
20 25 30
Ile Ser Ile Thr Ile Ile Ile Ser Ala Met Pro Thr Glu Ser Ile Ser
35 40 45
Ile Asp Asp Pro Tyr Phe Ser Thr Leu Cys Asn Thr Asn Pro Ser Ile
50 55 60
Thr Leu Ile His Leu Pro Gln Val Ser Leu Pro Pro Asn Thr Ser Phe
65 70 75 80
Ser Pro Leu Asp Phe Val Ala Ser Phe Phe Glu Leu Pro Glu Leu Asn
85 90 95
Asn Thr Asn Leu His Gln Thr Leu Leu Asn Leu Ser Lys Ser Ser Asn
100 105 110
Ile Lys Ala Phe Ile Ile Asp Phe Phe Cys Ser Ala Ala Phe Glu Phe
115 120 125
Val Ser Ser Arg His Asn Ile Pro Ile Tyr Phe Phe Tyr Thr Thr Cys
130 135 140
Ala Ser Gly Leu Ser Met Phe Leu His Leu Pro Ile Leu Asp Lys Ile
145 150 155 160
Ile Thr Lys Ser Leu Lys Asp Leu Asp Ile Ile Ile Asp Leu Pro Gly
165 170 175
Ile Pro Lys Ile Pro Ser Lys Glu Leu Pro Pro Ala Ile Ser Asp Arg
180 185 190
Ser His Arg Val Tyr Gln Tyr Leu Val Asp Thr Ala Lys Leu Met Ile
195 200 205
Lys Ser Ala Gly Leu Ile Ile Asn Thr Phe Glu Leu Leu Glu Arg Lys
210 215 220
Ala Leu Gln Ala Ile Gln Glu Gly Lys Cys Gly Ala Pro Asp Glu Pro
225 230 235 240
Val Pro Pro Leu Phe Cys Val Gly Pro Leu Leu Thr Thr Ser Glu Ser
245 250 255
Lys Ser Glu His Glu Cys Leu Thr Trp Leu Asp Ser Gln Pro Thr Arg
260 265 270
Ser Val Leu Phe Leu Cys Phe Gly Ser Met Gly Val Phe Asn Ser Arg
275 280 285
Gln Leu Arg Glu Thr Ala Ile Gly Leu Glu Lys Ser Gly Val Arg Phe
290 295 300
Leu Trp Val Val Arg Pro Pro Leu Ala Asp Ser Gln Thr Gln Ala Gly
305 310 315 320
Arg Ser Ser Thr Pro Asn Glu Pro Cys Leu Asp Leu Leu Leu Pro Glu
325 330 335
Gly Phe Leu Glu Arg Thr Lys Asp Arg Gly Phe Leu Val Asn Ser Trp
340 345 350
Ala Pro Gln Val Glu Ile Leu Asn His Gly Ser Val Gly Gly Phe Val
355 360 365
Thr His Cys Gly Trp Asn Ser Val Leu Glu Ala Leu Cys Ala Gly Val
370 375 380
Pro Met Val Ala Trp Pro Leu Tyr Ala Glu Gln Arg Met Asn Arg Ile
385 390 395 400
Phe Leu Val Glu Glu Met Lys Val Ala Leu Ala Phe Arg Glu Ala Gly
405 410 415
Asp Asp His Phe Val Asn Ala Ala Glu Leu Glu Glu Arg Val Ile Glu
420 425 430
Leu Met Asn Ser Lys Lys Gly Glu Ala Val Arg Glu Arg Val Leu Lys
435 440 445
Leu Arg Glu Asp Ala Val Val Ala Lys Ser Asp Gly Gly Ser Ser Cys
450 455 460
Ile Ala Met Ala Lys Leu Val Asp Cys Phe Lys Lys Gly
465 470 475
<210> 102
<211> 1434
<212> DNA
<213> 毛果杨
<400> 102
atggaagata ccattgttct gtatccgagt cctggtcgtg gtcacctgtt tagcatggtt 60
gaactgggta aacaaatcct ggaacatcat ccgagcatta gcattaccat tattatcagc 120
gcaatgccga ccgaaagcat cagcattgat gatccgtatt ttagcaccct gtgtaatacc 180
aatccgagta ttaccctgat tcatctgccg caggttagcc tgcctccgaa taccagcttt 240
agtccgctgg attttgttgc cagctttttt gaactgccgg aactgaataa tacgaatctg 300
catcagaccc tgctgaatct gagcaaaagc agcaacatta aagccttcat catcgacttt 360
ttttgcagcg cagcatttga atttgttagc agccgtcata acatcccgat ctattttttc 420
tataccacct gtgcaagcgg tctgagcatg tttctgcatc tgccgattct ggataaaatc 480
attaccaaaa gcctgaagga tctggatatt atcattgatc tgcctggcat tccgaaaatt 540
ccgagcaaag aactgcctcc ggcaattagc gatcgtagcc atcgtgttta tcagtatctg 600
gttgataccg ccaaactgat gattaaaagc gcaggtctga ttatcaacac ctttgagctg 660
ctggaacgta aagcactgca ggcaattcaa gagggtaaat gtggtgcacc ggatgaaccg 720
gtgcctccgc tgttttgtgt tggtccgctg ctgaccacca gtgaaagcaa aagcgaacat 780
gaatgtctga cctggctgga tagccagccg acacgtagcg ttctgtttct gtgttttggt 840
agcatgggtg tgtttaatag ccgtcagctg cgtgaaaccg caattggtct ggaaaaaagc 900
ggtgttcgtt ttctgtgggt tgttcgtccg cctctggcag atagtcagac ccaggcaggt 960
cgtagcagca ccccgaatga accgtgtctg gatctgctgc tgccggaagg ttttctggaa 1020
cgcaccaaag atcgtggctt tctggttaat agctgggcac cgcaggttga aattctgaat 1080
catggtagcg ttggtggttt tgttacccat tgtggttgga atagcgtgct ggaagcactg 1140
tgtgccggtg ttccgatggt tgcatggcct ctgtatgcag aacagcgtat gaatcgtatt 1200
tttctggtgg aagaaatgaa agttgcactg gcatttcgtg aagccggtga tgatcatttt 1260
gttaatgcag cagaactgga agaacgtgtg attgaactga tgaatagcaa aaaaggtgaa 1320
gccgttcgtg aacgtgttct gaaactgcgt gaagatgcag ttgttgcaaa aagtgatggt 1380
ggtagcagtt gtattgcaat ggcaaaactg gttgactgct ttaaaaaggg ctaa 1434
<210> 103
<211> 467
<212> PRT
<213> H. annuus
<400> 103
Met Glu Ser Ser Thr Val Val Met Tyr Pro Ser Pro Gly Ile Gly His
1 5 10 15
Leu Val Ser Met Val Glu Leu Gly Lys Leu Ile His Thr His His Pro
20 25 30
Ser Leu Ser Val Ile Ile Leu Ile Leu Thr Ala Pro Tyr Glu Thr Gly
35 40 45
Ala Thr Gly Lys Tyr Ile Asn Thr Val Ser Ala Thr Thr Pro Ala Ile
50 55 60
Thr Phe His His Leu Pro Ala Ile Ala Leu Pro Pro Asp Phe Ser Ser
65 70 75 80
Glu Phe Ile Asp Leu Ala Phe Gly Leu Pro Glu Leu Tyr Asn Ser Val
85 90 95
Val His Asn Thr Leu Val Ala Ile Ser Gln Lys Ser Thr Ile Lys Ala
100 105 110
Val Ile Leu Asp Phe Phe Ser Asn Ala Ala Phe Gln Val Ser Thr Asn
115 120 125
Leu Ser Leu Pro Thr Tyr Tyr Phe Phe Thr Ser Gly Thr Phe Gly Leu
130 135 140
Cys Ala Phe Leu Tyr Leu Thr Thr Leu His Lys Thr Thr Ser Lys Ser
145 150 155 160
Ile Lys Asp Leu Asn Thr Leu Leu Asp Phe Pro Gly Val Pro Pro Ile
165 170 175
His Ser Ser His Met Pro Thr Ala Ile Phe Asp Arg Glu Ser Asn Ser
180 185 190
Tyr Lys Asn Phe Met Lys Thr Ser Asn Asn Met Ala Lys Cys Ser Gly
195 200 205
Ile Ile Val Asn Ser Phe Leu Glu Leu Glu Glu Arg Ala Val Ala Thr
210 215 220
Leu Arg Asp Gly Lys Cys Ile Thr Asp Gly Pro Thr Pro Pro Ile Tyr
225 230 235 240
Phe Ile Gly Pro Leu Ile Ala Ser Gly Ser Gln Val Asp Pro Asn Glu
245 250 255
Asn Glu Cys Leu Lys Trp Leu Lys Thr Gln Pro Ser Lys Ser Val Val
260 265 270
Phe Leu Cys Phe Gly Ser Met Gly Val Phe Glu Lys Glu Gln Leu Lys
275 280 285
Glu Ile Ala Val Gly Leu Glu Arg Ser Gly Gln Arg Phe Leu Trp Val
290 295 300
Val Arg Asn Pro Pro Leu Glu Ser Ser Ser Gly Ala Lys Glu Phe Glu
305 310 315 320
Leu Asp Asp Ile Leu Pro Glu Gly Phe Leu Thr Arg Thr Lys Asp Lys
325 330 335
Gly Leu Val Val Lys Asn Trp Ala Pro Gln Pro Ala Ile Leu Gly His
340 345 350
Glu Ser Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Ser Leu
355 360 365
Glu Ala Val Val Ser Gly Val Pro Met Val Ala Trp Pro Leu Tyr Ala
370 375 380
Glu Gln Gln Met Asn Arg Val Tyr Leu Val Glu Glu Ile Lys Val Ala
385 390 395 400
Leu Trp Leu Arg Met Ser Ala Asp Gly Phe Val Gly Ala Glu Ala Val
405 410 415
Glu Glu Thr Val Arg Lys Leu Met Glu Gly Glu Glu Gly Arg Ala Val
420 425 430
Arg Glu Gln Ile Leu Glu Met Ser Gly Gly Ala Lys Ala Ala Val Glu
435 440 445
Asp Gly Gly Ser Ser Arg Leu Asp Phe Leu Lys Leu Thr Arg Pro Trp
450 455 460
Thr Asp Gln
465
<210> 104
<211> 1404
<212> DNA
<213> 向日葵
<400> 104
atggaaagca gcaccgttgt tatgtatccg agtcctggta ttggtcatct ggttagcatg 60
gttgaactgg gtaaactgat tcatacccat catccgagcc tgagcgttat tattctgatt 120
ctgaccgcac cgtatgaaac cggtgcaacc ggcaaatata tcaataccgt tagcgcaacc 180
acaccggcaa ttacctttca tcatctgcct gcaattgccc tgcctccgga ttttagcagc 240
gaatttattg atctggcatt tggtctgccg gaactgtata atagcgttgt tcataatacc 300
ctggttgcca ttagccagaa aagcaccatt aaagcagtta tcctggattt ctttagcaac 360
gcagcatttc aggttagcac caatctgagc ctgccgacct attatttctt taccagcggc 420
acctttggtc tgtgtgcatt tctgtatctg accacactgc ataaaaccac gagcaaaagc 480
attaaagatc tgaataccct gctggatttt ccgggtgttc cgcctattca tagcagccat 540
atgccgaccg caatttttga tcgtgaaagc aacagctaca aaaactttat gaaaaccagc 600
aacaacatgg ccaaatgcag cggtattatt gtgaatagct ttctggaact ggaagaacgt 660
gcagttgcaa ccctgcgtga tggtaaatgt attaccgatg gtccgacacc tccgatttat 720
ttcattggtc cgctgattgc aagcggtagc caggttgatc cgaatgaaaa tgaatgtctg 780
aaatggctga aaacccagcc gagcaaatca gttgtttttc tgtgttttgg tagcatgggc 840
gtgtttgaaa aagaacagct gaaagaaatt gccgttggtc tggaacgtag cggtcagcgt 900
tttctgtggg ttgttcgtaa tccgcctctg gaaagctcaa gcggtgcaaa agaatttgaa 960
ctggatgata tcctgccgga aggttttctg acccgtacca aagataaagg tctggttgtg 1020
aaaaattggg caccgcagcc tgccattctg ggtcatgaaa gcgttggtgg ttttgttagc 1080
cattgtggtt ggaatagcag cctggaagca gttgttagcg gtgttccgat ggttgcatgg 1140
cctctgtatg cagaacagca gatgaatcgt gtttatctgg tggaagaaat taaagttgca 1200
ctgtggctgc gtatgagcgc agatggtttt gtgggtgcag aagccgttga agaaaccgtt 1260
cgcaaactga tggaaggtga agagggtcgt gcagttcgtg agcagattct ggaaatgagc 1320
ggtggtgcca aagcagcagt tgaagatggt ggtagcagcc gtctggattt cctgaaactg 1380
acccgtccgt ggaccgatca gtaa 1404
<210> 105
<211> 458
<212> PRT
<213> 甜叶菊(S. rebaudiana)
<400> 105
Met Glu Asn Lys Thr Glu Thr Thr Val Arg Arg Arg Arg Arg Ile Ile
1 5 10 15
Leu Phe Pro Val Pro Phe Gln Gly His Ile Asn Pro Ile Leu Gln Leu
20 25 30
Ala Asn Val Leu Tyr Ser Lys Gly Phe Ser Ile Thr Ile Phe His Thr
35 40 45
Asn Phe Asn Lys Pro Lys Thr Ser Asn Tyr Pro His Phe Thr Phe Arg
50 55 60
Phe Ile Leu Asp Asn Asp Pro Gln Asp Glu Arg Ile Ser Asn Leu Pro
65 70 75 80
Thr His Gly Pro Leu Ala Gly Met Arg Ile Pro Ile Ile Asn Glu His
85 90 95
Gly Ala Asp Glu Leu Arg Arg Glu Leu Glu Leu Leu Met Leu Ala Ser
100 105 110
Glu Glu Asp Glu Glu Val Ser Cys Leu Ile Thr Asp Ala Leu Trp Tyr
115 120 125
Phe Ala Gln Ser Val Ala Asp Ser Leu Asn Leu Arg Arg Leu Val Leu
130 135 140
Met Thr Ser Ser Leu Phe Asn Phe His Ala His Val Ser Leu Pro Gln
145 150 155 160
Phe Asp Glu Leu Gly Tyr Leu Asp Pro Asp Asp Lys Thr Arg Leu Glu
165 170 175
Glu Gln Ala Ser Gly Phe Pro Met Leu Lys Val Lys Asp Ile Lys Ser
180 185 190
Ala Tyr Ser Asn Trp Gln Ile Leu Lys Glu Ile Leu Gly Lys Met Ile
195 200 205
Lys Gln Thr Lys Ala Ser Ser Gly Val Ile Trp Asn Ser Phe Lys Glu
210 215 220
Leu Glu Glu Ser Glu Leu Glu Thr Val Ile Arg Glu Ile Pro Ala Pro
225 230 235 240
Ser Phe Leu Ile Pro Leu Pro Lys His Leu Thr Ala Ser Ser Ser Ser
245 250 255
Leu Leu Asp His Asp Arg Thr Val Phe Gln Trp Leu Asp Gln Gln Pro
260 265 270
Pro Ser Ser Val Leu Tyr Val Ser Phe Gly Ser Thr Ser Glu Val Asp
275 280 285
Glu Lys Asp Phe Leu Glu Ile Ala Arg Gly Leu Val Asp Ser Lys Gln
290 295 300
Ser Phe Leu Trp Val Val Arg Pro Gly Phe Val Lys Gly Ser Thr Trp
305 310 315 320
Val Glu Pro Leu Pro Asp Gly Phe Leu Gly Glu Arg Gly Arg Ile Val
325 330 335
Lys Trp Val Pro Gln Gln Glu Val Leu Ala His Gly Ala Ile Gly Ala
340 345 350
Phe Trp Thr His Ser Gly Trp Asn Ser Thr Leu Glu Ser Val Cys Glu
355 360 365
Gly Val Pro Met Ile Phe Ser Asp Phe Gly Leu Asp Gln Pro Leu Asn
370 375 380
Ala Arg Tyr Met Ser Asp Val Leu Lys Val Gly Val Tyr Leu Glu Asn
385 390 395 400
Gly Trp Glu Arg Gly Glu Ile Ala Asn Ala Ile Arg Arg Val Met Val
405 410 415
Asp Glu Glu Gly Glu Tyr Ile Arg Gln Asn Ala Arg Val Leu Lys Gln
420 425 430
Lys Ala Asp Val Ser Leu Met Lys Gly Gly Ser Ser Tyr Glu Ser Leu
435 440 445
Glu Ser Leu Val Ser Tyr Ile Ser Ser Leu
450 455
<210> 106
<211> 1377
<212> DNA
<213> 甜叶菊
<400> 106
atggaaaaca aaaccgaaac caccgtgcgt cgtcgtcgcc gtattattct gtttccggtt 60
ccgtttcagg gtcatattaa tccgattctg cagctggcaa atgtgctgta tagcaaaggt 120
tttagcatca ccatctttca caccaacttc aacaaaccga aaaccagcaa ttatccgcat 180
tttacctttc gctttatcct ggataatgat ccgcaggatg aacgtattag caatctgccg 240
acacatggtc cgctggcagg tatgcgtatt ccgattatta acgaacatgg tgcagatgaa 300
ctgcgtcgtg aactggaact gctgatgctg gcaagcgaag aagatgaaga agttagctgt 360
ctgattaccg atgcactgtg gtattttgca cagagcgttg cagatagcct gaatctgcgt 420
cgcctggttc tgatgaccag cagcctgttt aactttcatg cacatgttag cctgccgcag 480
tttgatgaac tgggttatct ggatccggat gataaaaccc gtctggaaga acaggcaagc 540
ggttttccga tgctgaaagt gaaagatatc aaaagcgcat atagcaactg gcagatcctg 600
aaagaaattc tgggcaaaat gatcaaacag accaaagcaa gcagcggtgt tatttggaat 660
agctttaaag aactggaaga gagcgaactg gaaaccgtta ttcgtgaaat tccggcaccg 720
agctttctga ttccgctgcc gaaacatctg accgcaagca gcagcagtct gctggatcac 780
gatcgtaccg tttttcagtg gctggatcag cagcctccga gcagcgttct gtatgttagc 840
tttggtagca ccagcgaagt tgatgaaaaa gactttctgg aaattgcacg tggtctggtt 900
gatagcaaac agagttttct gtgggttgtt cgtccgggtt ttgttaaagg tagcacctgg 960
gttgaaccgc tgccggatgg ttttctgggt gaacgtggtc gtattgttaa atgggttccg 1020
cagcaagagg ttctggcaca tggtgccatt ggtgcatttt ggacccatag cggttggaat 1080
agtaccctgg aaagcgtttg tgaaggtgtt ccgatgattt ttagcgattt tggtctggat 1140
caaccgctga atgcacgtta tatgagtgat gttctgaaag tgggtgtgta tctggaaaat 1200
ggttgggaac gtggtgaaat tgcaaatgca attcgtcgtg ttatggttga tgaagagggt 1260
gaatatatcc gtcagaatgc ccgtgtgctg aaacagaaag cagatgtgag cctgatgaaa 1320
ggtggtagca gctatgaaag cctggaaagt ctggttagct atatcagctc actgtaa 1377
<210> 107
<211> 495
<212> PRT
<213> A. thaliana
<400> 107
Met Val Ser Glu Thr Thr Lys Ser Ser Pro Leu His Phe Val Leu Phe
1 5 10 15
Pro Phe Met Ala Gln Gly His Met Ile Pro Met Val Asp Ile Ala Arg
20 25 30
Leu Leu Ala Gln Arg Gly Val Ile Ile Thr Ile Val Thr Thr Pro His
35 40 45
Asn Ala Ala Arg Phe Lys Asn Val Leu Asn Arg Ala Ile Glu Ser Gly
50 55 60
Leu Pro Ile Asn Leu Val Gln Val Lys Phe Pro Tyr Leu Glu Ala Gly
65 70 75 80
Leu Gln Glu Gly Gln Glu Asn Ile Asp Ser Leu Asp Thr Met Glu Arg
85 90 95
Met Ile Pro Phe Phe Lys Ala Val Asn Phe Leu Glu Glu Pro Val Gln
100 105 110
Lys Leu Ile Glu Glu Met Asn Pro Arg Pro Ser Cys Leu Ile Ser Asp
115 120 125
Phe Cys Leu Pro Tyr Thr Ser Lys Ile Ala Lys Lys Phe Asn Ile Pro
130 135 140
Lys Ile Leu Phe His Gly Met Gly Cys Phe Cys Leu Leu Cys Met His
145 150 155 160
Val Leu Arg Lys Asn Arg Glu Ile Leu Asp Asn Leu Lys Ser Asp Lys
165 170 175
Glu Leu Phe Thr Val Pro Asp Phe Pro Asp Arg Val Glu Phe Thr Arg
180 185 190
Thr Gln Val Pro Val Glu Thr Tyr Val Pro Ala Gly Asp Trp Lys Asp
195 200 205
Ile Phe Asp Gly Met Val Glu Ala Asn Glu Thr Ser Tyr Gly Val Ile
210 215 220
Val Asn Ser Phe Gln Glu Leu Glu Pro Ala Tyr Ala Lys Asp Tyr Lys
225 230 235 240
Glu Val Arg Ser Gly Lys Ala Trp Thr Ile Gly Pro Val Ser Leu Cys
245 250 255
Asn Lys Val Gly Ala Asp Lys Ala Glu Arg Gly Asn Lys Ser Asp Ile
260 265 270
Asp Gln Asp Glu Cys Leu Lys Trp Leu Asp Ser Lys Lys His Gly Ser
275 280 285
Val Leu Tyr Val Cys Leu Gly Ser Ile Cys Asn Leu Pro Leu Ser Gln
290 295 300
Leu Lys Glu Leu Gly Leu Gly Leu Glu Glu Ser Gln Arg Pro Phe Ile
305 310 315 320
Trp Val Ile Arg Gly Trp Glu Lys Tyr Lys Glu Leu Val Glu Trp Phe
325 330 335
Ser Glu Ser Gly Phe Glu Asp Arg Ile Gln Asp Arg Gly Leu Leu Ile
340 345 350
Lys Gly Trp Ser Pro Gln Met Leu Ile Leu Ser His Pro Ser Val Gly
355 360 365
Gly Phe Leu Thr His Cys Gly Trp Asn Ser Thr Leu Glu Gly Ile Thr
370 375 380
Ala Gly Leu Pro Leu Leu Thr Trp Pro Leu Phe Ala Asp Gln Phe Cys
385 390 395 400
Asn Glu Lys Leu Val Val Glu Val Leu Lys Ala Gly Val Arg Ser Gly
405 410 415
Val Glu Gln Pro Met Lys Trp Gly Glu Glu Glu Lys Ile Gly Val Leu
420 425 430
Val Asp Lys Glu Gly Val Lys Lys Ala Val Glu Glu Leu Met Gly Glu
435 440 445
Ser Asp Asp Ala Lys Glu Arg Arg Arg Arg Ala Lys Glu Leu Gly Asp
450 455 460
Ser Ala His Lys Ala Val Glu Glu Gly Gly Ser Ser His Ser Asn Ile
465 470 475 480
Ser Phe Leu Leu Gln Asp Ile Met Glu Leu Ala Glu Pro Asn Asn
485 490 495
<210> 108
<211> 1488
<212> DNA
<213> 拟南芥
<400> 108
atggttagcg aaaccaccaa aagcagtccg ctgcattttg ttctgtttcc gtttatggca 60
cagggtcata tgattccgat ggttgatatt gcacgtctgc tggcacagcg tggtgtgatt 120
attaccattg ttaccacacc gcataatgca gcacgcttta aaaacgttct gaatcgtgca 180
attgaaagcg gtctgccgat taatctggtt caggttaaat ttccgtatct ggaagcaggt 240
ctgcaagaag gtcaagaaaa tattgatagc ctggatacca tggaacgcat gattccgttt 300
ttcaaagccg tgaattttct ggaagaaccg gtgcagaaac tgatcgaaga aatgaatccg 360
cgtccgagct gtctgattag cgatttttgt ctgccgtata ccagcaaaat cgccaaaaaa 420
ttcaacatcc cgaaaatcct gtttcatggt atgggttgtt tttgcctgct gtgtatgcat 480
gttctgcgta aaaatcgtga aatcctggat aacctgaaaa gcgataaaga actgtttacc 540
gttccggatt ttccggatcg tgtggaattt acccgtacac aggttccggt tgaaacctat 600
gttccggcag gcgattggaa agatattttt gatggtatgg tggaagccaa cgaaaccagc 660
tatggtgtta ttgtgaatag ctttcaagaa ctggaaccgg catatgcgaa agattacaaa 720
gaagttcgta gcggtaaagc atggaccatt ggtccggtta gcctgtgtaa taaagttggt 780
gcagataaag cagaacgcgg taataaaagt gatatcgatc aggatgaatg cctgaaatgg 840
ctggatagca aaaaacatgg tagcgttctg tatgtttgtc tgggtagcat ttgcaatctg 900
ccgctgagcc agctgaaaga attaggtctg ggtttagaag aaagccagcg tccgtttatt 960
tgggttattc gtggttggga gaaatacaaa gaactggttg aatggttttc cgaaagcggt 1020
tttgaagatc gtattcagga tcgtggcctg ctgattaaag gttggagtcc gcagatgctg 1080
attctgagcc atccgagcgt tggtggcttt ctgacccatt gtggttggaa tagcaccctg 1140
gaaggtatta cagctggcct gccgctgctg acctggcctc tgtttgcaga tcagttttgt 1200
aatgaaaaac tggtggtgga agttctgaaa gccggtgtgc gtagcggtgt tgaacagccg 1260
atgaaatggg gtgaagaaga aaaaattggc gtcctggttg ataaagaagg tgttaaaaaa 1320
gccgtggaag aactgatggg tgaaagtgat gatgcaaaag aacgtcgtcg tcgtgcaaaa 1380
gagctgggcg atagcgcaca taaagcagtt gaagaaggtg gtagcagcca tagcaatatt 1440
agctttctgc tgcaggatat tatggaactg gcagaaccga ataactaa 1488
<210> 109
<211> 467
<212> PRT
<213> 拟南芥
<400> 109
Met Arg Asn Val Glu Leu Ile Phe Ile Pro Thr Pro Thr Val Gly His
1 5 10 15
Leu Val Pro Phe Leu Glu Phe Ala Arg Arg Leu Ile Glu Gln Asp Asp
20 25 30
Arg Ile Arg Ile Thr Ile Leu Leu Met Lys Leu Gln Gly Gln Ser His
35 40 45
Leu Asp Thr Tyr Val Lys Ser Ile Ala Ser Ser Gln Pro Phe Val Arg
50 55 60
Phe Ile Asp Val Pro Glu Leu Glu Glu Lys Pro Thr Leu Gly Ser Thr
65 70 75 80
Gln Ser Val Glu Ala Tyr Val Tyr Asp Val Ile Glu Arg Asn Ile Pro
85 90 95
Leu Val Arg Asn Ile Val Met Asp Ile Leu Thr Ser Leu Ala Leu Asp
100 105 110
Gly Val Lys Val Lys Gly Leu Val Val Asp Phe Phe Cys Leu Pro Met
115 120 125
Ile Asp Val Ala Lys Asp Ile Ser Leu Pro Phe Tyr Val Phe Leu Thr
130 135 140
Thr Asn Ser Gly Phe Leu Ala Met Met Gln Tyr Leu Ala Asp Arg His
145 150 155 160
Ser Arg Asp Thr Ser Val Phe Val Arg Asn Ser Glu Glu Met Leu Ser
165 170 175
Ile Pro Gly Phe Val Asn Pro Val Pro Ala Asn Val Leu Pro Ser Ala
180 185 190
Leu Phe Val Glu Asp Gly Tyr Asp Ala Tyr Val Lys Leu Ala Ile Leu
195 200 205
Phe Thr Lys Ala Asn Gly Ile Leu Val Asn Ser Ser Phe Asp Ile Glu
210 215 220
Pro Tyr Ser Val Asn His Phe Leu Gln Glu Gln Asn Tyr Pro Ser Val
225 230 235 240
Tyr Ala Val Gly Pro Ile Phe Asp Leu Lys Ala Gln Pro His Pro Glu
245 250 255
Gln Asp Leu Thr Arg Arg Asp Glu Leu Met Lys Trp Leu Asp Asp Gln
260 265 270
Pro Glu Ala Ser Val Val Phe Leu Cys Phe Gly Ser Met Ala Arg Leu
275 280 285
Arg Gly Ser Leu Val Lys Glu Ile Ala His Gly Leu Glu Leu Cys Gln
290 295 300
Tyr Arg Phe Leu Trp Ser Leu Arg Lys Glu Glu Val Thr Lys Asp Asp
305 310 315 320
Leu Pro Glu Gly Phe Leu Asp Arg Val Asp Gly Arg Gly Met Ile Cys
325 330 335
Gly Trp Ser Pro Gln Val Glu Ile Leu Ala His Lys Ala Val Gly Gly
340 345 350
Phe Val Ser His Cys Gly Trp Asn Ser Ile Val Glu Ser Leu Trp Phe
355 360 365
Gly Val Pro Ile Val Thr Trp Pro Met Tyr Ala Glu Gln Gln Leu Asn
370 375 380
Ala Phe Leu Met Val Lys Glu Leu Lys Leu Ala Val Glu Leu Lys Leu
385 390 395 400
Asp Tyr Arg Val His Ser Asp Glu Ile Val Asn Ala Asn Glu Ile Glu
405 410 415
Thr Ala Ile Arg Tyr Val Met Asp Thr Asp Asn Asn Val Val Arg Lys
420 425 430
Arg Val Met Asp Ile Ser Gln Met Ile Gln Arg Ala Thr Lys Asn Gly
435 440 445
Gly Ser Ser Phe Ala Ala Ile Glu Lys Phe Ile Tyr Asp Val Ile Gly
450 455 460
Ile Lys Pro
465
<210> 110
<211> 1404
<212> DNA
<213> 拟南芥
<400> 110
atgcgtaatg tggaactgat ttttatcccg acaccgaccg ttggtcatct ggttccgttt 60
ctggaatttg cacgtcgtct gattgaacag gatgatcgta ttcgtattac catcctgctg 120
atgaaactgc agggtcagag ccatctggat acctatgtta aaagcattgc aagcagccag 180
ccgtttgttc gttttattga tgtgccggaa ctggaagaaa aaccgacact gggtagcacc 240
cagagcgttg aagcatatgt ttatgatgtg attgaacgca atattccgct ggtgcgtaat 300
attgttatgg atattctgac cagcctggca ctggatggtg ttaaagttaa aggtctggtt 360
gtggattttt tctgcctgcc gatgattgat gttgccaaag atattagcct gccgttttat 420
gtttttctga ccaccaatag cggttttctg gcaatgatgc agtatctggc agatcgtcat 480
agccgtgata ccagcgtttt tgttcgtaat agcgaagaaa tgctgagcat tccgggtttt 540
gttaatccgg ttccggcaaa tgttctgccg agcgcactgt ttgttgaaga tggttatgat 600
gcgtatgtta aactggccat cctgtttacc aaagccaatg gtattctggt gaatagcagc 660
tttgatatcg aaccgtatag cgtgaatcac tttctgcaag aacagaatta tccgagcgtt 720
tatgcagttg gtccgatctt tgatctgaaa gcacagccgc atccggaaca ggatctgacc 780
cgtcgtgatg aactgatgaa atggctggat gatcagccgg aagcaagcgt tgtgtttctg 840
tgttttggta gcatggcacg tctgcgtggt agcctggtta aagaaattgc acatggtctg 900
gaactgtgcc agtatcgttt tctgtggtca ctgcgtaaag aagaagttac caaagacgac 960
ctgccggaag gctttctgga tcgtgttgat ggtcgtggta tgatttgtgg ttggagtccg 1020
caggttgaaa ttctggcaca taaagcagtt ggtggttttg tgagccattg cggttggaat 1080
agcattgttg aaagcctgtg gtttggtgtt ccgattgtta cctggccgat gtatgcagaa 1140
cagcagctga atgcatttct gatggtgaaa gaactgaaac tggcagttga actgaagctg 1200
gattatcgtg ttcattccga tgaaattgtg aacgccaatg aaattgaaac cgccattcgt 1260
tatgtgatgg ataccgataa caatgttgtg cgtaaacgtg tcatggatat cagccagatg 1320
attcagcgtg caaccaaaaa tggtggtagc agttttgcag ccatcgagaa atttatctat 1380
gacgtgattg gcatcaagcc gtaa 1404
<210> 111
<211> 480
<212> PRT
<213> 拟南芥
<400> 111
Met Glu Glu Ser Lys Thr Pro His Val Ala Ile Ile Pro Ser Pro Gly
1 5 10 15
Met Gly His Leu Ile Pro Leu Val Glu Phe Ala Lys Arg Leu Val His
20 25 30
Leu His Gly Leu Thr Val Thr Phe Val Ile Ala Gly Glu Gly Pro Pro
35 40 45
Ser Lys Ala Gln Arg Thr Val Leu Asp Ser Leu Pro Ser Ser Ile Ser
50 55 60
Ser Val Phe Leu Pro Pro Val Asp Leu Thr Asp Leu Ser Ser Ser Thr
65 70 75 80
Arg Ile Glu Ser Arg Ile Ser Leu Thr Val Thr Arg Ser Asn Pro Glu
85 90 95
Leu Arg Lys Val Phe Asp Ser Phe Val Glu Gly Gly Arg Leu Pro Thr
100 105 110
Ala Leu Val Val Asp Leu Phe Gly Thr Asp Ala Phe Asp Val Ala Val
115 120 125
Glu Phe His Val Pro Pro Tyr Ile Phe Tyr Pro Thr Thr Ala Asn Val
130 135 140
Leu Ser Phe Phe Leu His Leu Pro Lys Leu Asp Glu Thr Val Ser Cys
145 150 155 160
Glu Phe Arg Glu Leu Thr Glu Pro Leu Met Leu Pro Gly Cys Val Pro
165 170 175
Val Ala Gly Lys Asp Phe Leu Asp Pro Ala Gln Asp Arg Lys Asp Asp
180 185 190
Ala Tyr Lys Trp Leu Leu His Asn Thr Lys Arg Tyr Lys Glu Ala Glu
195 200 205
Gly Ile Leu Val Asn Thr Phe Phe Glu Leu Glu Pro Asn Ala Ile Lys
210 215 220
Ala Leu Gln Glu Pro Gly Leu Asp Lys Pro Pro Val Tyr Pro Val Gly
225 230 235 240
Pro Leu Val Asn Ile Gly Lys Gln Glu Ala Lys Gln Thr Glu Glu Ser
245 250 255
Glu Cys Leu Lys Trp Leu Asp Asn Gln Pro Leu Gly Ser Val Leu Tyr
260 265 270
Val Ser Phe Gly Ser Gly Gly Thr Leu Thr Cys Glu Gln Leu Asn Glu
275 280 285
Leu Ala Leu Gly Leu Ala Asp Ser Glu Gln Arg Phe Leu Trp Val Ile
290 295 300
Arg Ser Pro Ser Gly Ile Ala Asn Ser Ser Tyr Phe Asp Ser His Ser
305 310 315 320
Gln Thr Asp Pro Leu Thr Phe Leu Pro Pro Gly Phe Leu Glu Arg Thr
325 330 335
Lys Lys Arg Gly Phe Val Ile Pro Phe Trp Ala Pro Gln Ala Gln Val
340 345 350
Leu Ala His Pro Ser Thr Gly Gly Phe Leu Thr His Cys Gly Trp Asn
355 360 365
Ser Thr Leu Glu Ser Val Val Ser Gly Ile Pro Leu Ile Ala Trp Pro
370 375 380
Leu Tyr Ala Glu Gln Lys Met Asn Ala Val Leu Leu Ser Glu Asp Ile
385 390 395 400
Arg Ala Ala Leu Arg Pro Arg Ala Gly Asp Asp Gly Leu Val Arg Arg
405 410 415
Glu Glu Val Ala Arg Val Val Lys Gly Leu Met Glu Gly Glu Glu Gly
420 425 430
Lys Gly Val Arg Asn Lys Met Lys Glu Leu Lys Glu Ala Ala Cys Arg
435 440 445
Val Leu Lys Asp Asp Gly Thr Ser Thr Lys Ala Leu Ser Leu Val Ala
450 455 460
Leu Lys Trp Lys Ala His Lys Lys Glu Leu Glu Gln Asn Gly Asn His
465 470 475 480
<210> 112
<211> 1443
<212> DNA
<213> 拟南芥
<400> 112
atggaagaaa gcaaaacacc gcatgttgca attattccga gtcctggtat gggtcatctg 60
attccgctgg ttgaatttgc aaaacgtctg gttcatctgc atggtctgac cgttaccttt 120
gttattgccg gtgaaggtcc gcctagcaaa gcacagcgta ccgttctgga tagcctgccg 180
agcagcatta gcagcgtttt tctgcctccg gttgatctga ccgatctgag cagcagcacc 240
cgtattgaaa gccgtattag cctgacagtt acccgtagca atccggaact gcgtaaagtt 300
tttgatagct ttgttgaagg tggtcgtctg ccgaccgcac tggttgttga cctgtttggc 360
accgatgcat ttgatgttgc agttgaattt catgtgcctc cgtatatctt ttatccgacc 420
accgcaaatg ttctgagctt ttttctgcat ctgccgaaac tggatgaaac cgttagctgt 480
gaatttcgtg aactgaccga accgctgatg ctgcctggtt gtgttccggt tgcaggtaaa 540
gattttctgg atccggcaca ggatcgtaaa gatgatgcat ataaatggct gctgcataac 600
accaaacgtt ataaagaagc agaaggcatt ctggtcaaca ccttttttga actggaaccg 660
aatgcaatta aagccctgca agaacctggt ctggataaac cgcctgttta tccggttggt 720
cctctggtta atattggtaa acaagaagcc aaacagaccg aagaaagcga atgtctgaaa 780
tggctggata atcagccgct gggtagcgtt ctgtatgtta gctttggtag cggtggcacc 840
ctgacctgtg aacagctgaa tgaactggca ctgggtttag cagatagcga acagcgtttt 900
ctgtgggtta ttcgtagccc gagcggtatt gcaaatagca gttattttga tagtcacagc 960
cagacagatc cgctgacctt tctgccaccg ggttttctgg aacgtaccaa aaaacgtggt 1020
tttgtgattc cgttttgggc accgcaggca caggttctgg cacatccgag caccggtggt 1080
tttctgaccc attgtggttg gaatagcacc ctggaaagcg ttgttagcgg tattccgctg 1140
attgcatggc ctctgtatgc agaacagaaa atgaatgcag ttctgctgag cgaagatatt 1200
cgtgcagcac tgcgtccgcg tgccggtgat gatggtctgg ttcgtcgtga agaagttgca 1260
cgcgttgtta aaggtctgat ggaaggtgaa gaaggtaaag gcgttcgcaa caaaatgaaa 1320
gaactgaaag aggcagcctg tcgcgttctg aaagatgacg gcaccagcac caaagcactg 1380
agcctggttg cactgaaatg gaaagcacat aaaaaagagc tggaacagaa cggcaaccac 1440
taa 1443
<210> 113
<211> 474
<212> PRT
<213> 甜叶菊
<400> 113
Met Ser Thr Ser Glu Leu Val Phe Ile Pro Ser Pro Gly Ala Gly His
1 5 10 15
Leu Pro Pro Thr Val Glu Leu Ala Lys Leu Leu Leu His Arg Asp Gln
20 25 30
Arg Leu Ser Val Thr Ile Ile Val Met Asn Leu Trp Leu Gly Pro Lys
35 40 45
His Asn Thr Glu Ala Arg Pro Cys Val Pro Ser Leu Arg Phe Val Asp
50 55 60
Ile Pro Cys Asp Glu Ser Thr Met Ala Leu Ile Ser Pro Asn Thr Phe
65 70 75 80
Ile Ser Ala Phe Val Glu His His Lys Pro Arg Val Arg Asp Ile Val
85 90 95
Arg Gly Ile Ile Glu Ser Asp Ser Val Arg Leu Ala Gly Phe Val Leu
100 105 110
Asp Met Phe Cys Met Pro Met Ser Asp Val Ala Asn Glu Phe Gly Val
115 120 125
Pro Ser Tyr Asn Tyr Phe Thr Ser Gly Ala Ala Thr Leu Gly Leu Met
130 135 140
Phe His Leu Gln Trp Lys Arg Asp His Glu Gly Tyr Asp Ala Thr Glu
145 150 155 160
Leu Lys Asn Ser Asp Thr Glu Leu Ser Val Pro Ser Tyr Val Asn Pro
165 170 175
Val Pro Ala Lys Val Leu Pro Glu Val Val Leu Asp Lys Glu Gly Gly
180 185 190
Ser Lys Met Phe Leu Asp Leu Ala Glu Arg Ile Arg Glu Ser Lys Gly
195 200 205
Ile Ile Val Asn Ser Cys Gln Ala Ile Glu Arg His Ala Leu Glu Tyr
210 215 220
Leu Ser Ser Asn Asn Asn Gly Ile Pro Pro Val Phe Pro Val Gly Pro
225 230 235 240
Ile Leu Asn Leu Glu Asn Lys Lys Asp Asp Ala Lys Thr Asp Glu Ile
245 250 255
Met Arg Trp Leu Asn Glu Gln Pro Glu Ser Ser Val Val Phe Leu Cys
260 265 270
Phe Gly Ser Met Gly Ser Phe Asn Glu Lys Gln Val Lys Glu Ile Ala
275 280 285
Val Ala Ile Glu Arg Ser Gly His Arg Phe Leu Trp Ser Leu Arg Arg
290 295 300
Pro Thr Pro Lys Glu Lys Ile Glu Phe Pro Lys Glu Tyr Glu Asn Leu
305 310 315 320
Glu Glu Val Leu Pro Glu Gly Phe Leu Lys Arg Thr Ser Ser Ile Gly
325 330 335
Lys Val Ile Gly Trp Ala Pro Gln Met Ala Val Leu Ser His Pro Ser
340 345 350
Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Thr Leu Glu Ser
355 360 365
Met Trp Cys Gly Val Pro Met Ala Ala Trp Pro Leu Tyr Ala Glu Gln
370 375 380
Thr Leu Asn Ala Phe Leu Leu Val Val Glu Leu Gly Leu Ala Ala Glu
385 390 395 400
Ile Arg Met Asp Tyr Arg Thr Asp Thr Lys Ala Gly Tyr Asp Gly Gly
405 410 415
Met Glu Val Thr Val Glu Glu Ile Glu Asp Gly Ile Arg Lys Leu Met
420 425 430
Ser Asp Gly Glu Ile Arg Asn Lys Val Lys Asp Val Lys Glu Lys Ser
435 440 445
Arg Ala Ala Val Val Glu Gly Gly Ser Ser Tyr Ala Ser Ile Gly Lys
450 455 460
Phe Ile Glu His Val Ser Asn Val Thr Ile
465 470
<210> 114
<211> 1425
<212> DNA
<213> 甜叶菊
<400> 114
atgagcacca gcgaactggt ttttattccg agtcctggtg caggtcatct gcctccgacc 60
gttgaactgg caaaactgct gctgcatcgt gatcagcgtc tgagcgttac cattattgtt 120
atgaatctgt ggctgggtcc gaaacataat accgaagcac gtccgtgtgt tccgagcctg 180
cgttttgttg atattccgtg tgatgaaagc accatggcac tgattagccc gaataccttt 240
attagcgcat ttgtggaaca tcataaaccg cgtgttcgtg atattgtgcg tggtattatt 300
gaaagcgata gcgttcgtct ggcaggtttt gttctggata tgttttgtat gccgatgagt 360
gatgtggcca atgaatttgg tgtgccgagc tataactatt ttaccagcgg tgcagcaacc 420
ctgggtctga tgtttcatct gcagtggaaa cgtgatcatg aaggttatga tgcaaccgaa 480
ctgaaaaata gcgataccga actgtcagtt ccgagctatg ttaatccggt tccggcaaaa 540
gttctgcctg aagttgtgct ggataaagaa ggtggtagca aaatgtttct ggatctggca 600
gaacgtattc gtgaaagcaa aggcattatt gtgaatagct gtcaggcaat tgaacgtcat 660
gcactggaat atctgagcag caataacaat ggtattccgc ctgtttttcc ggttggtccg 720
attctgaatc tggaaaacaa aaaagatgat gccaaaaccg atgaaattat gcgctggctg 780
aatgaacagc cggaaagcag cgttgttttt ctgtgttttg gtagcatggg cagctttaat 840
gagaaacagg ttaaagaaat tgccgtggcc attgaacgta gcggtcatcg ttttctgtgg 900
tcactgcgtc gtccgacacc gaaagaaaaa attgaatttc cgaaagaata tgagaacctg 960
gaagaagtgc tgccggaagg ttttctgaaa cgtaccagca gcattggtaa agttattggt 1020
tgggcaccgc agatggcagt tctgagccat ccgagcgttg gtggttttgt tagccattgt 1080
ggttggaata gcaccctgga aagcatgtgg tgtggtgttc cgatggcagc atggcctctg 1140
tatgcagaac agaccctgaa tgcatttctg ctggttgttg aattaggtct ggcagccgaa 1200
attcgtatgg attatcgtac cgataccaaa gcaggctatg atggtggtat ggaagttacc 1260
gttgaagaaa ttgaagatgg cattcgcaaa ctgatgtcag atggtgaaat tcgcaacaaa 1320
gtgaaggacg tgaaagagaa aagtcgcgca gcagttgttg aaggtggttc aagctatgca 1380
agtatcggca aattcatcga acatgttagc aacgtgacca tttaa 1425
<210> 115
<211> 462
<212> PRT
<213> 水稻
<400> 115
Met Asp Ser Gly Tyr Ser Ser Ser Tyr Ala Ala Ala Ala Gly Met His
1 5 10 15
Val Val Ile Cys Pro Trp Leu Ala Phe Gly His Leu Leu Pro Cys Leu
20 25 30
Asp Leu Ala Gln Arg Leu Ala Ser Arg Gly His Arg Val Ser Phe Val
35 40 45
Ser Thr Pro Arg Asn Ile Ser Arg Leu Pro Pro Val Arg Pro Ala Leu
50 55 60
Ala Pro Leu Val Ala Phe Val Ala Leu Pro Leu Pro Arg Val Glu Gly
65 70 75 80
Leu Pro Asp Gly Ala Glu Ser Thr Asn Asp Val Pro His Asp Arg Pro
85 90 95
Asp Met Val Glu Leu His Arg Arg Ala Phe Asp Gly Leu Ala Ala Pro
100 105 110
Phe Ser Glu Phe Leu Gly Thr Ala Cys Ala Asp Trp Val Ile Val Asp
115 120 125
Val Phe His His Trp Ala Ala Ala Ala Ala Leu Glu His Lys Val Pro
130 135 140
Cys Ala Met Met Leu Leu Gly Ser Ala His Met Ile Ala Ser Ile Ala
145 150 155 160
Asp Arg Arg Leu Glu Arg Ala Glu Thr Glu Ser Pro Ala Ala Ala Gly
165 170 175
Gln Gly Arg Pro Ala Ala Ala Pro Thr Phe Glu Val Ala Arg Met Lys
180 185 190
Leu Ile Arg Thr Lys Gly Ser Ser Gly Met Ser Leu Ala Glu Arg Phe
195 200 205
Ser Leu Thr Leu Ser Arg Ser Ser Leu Val Val Gly Arg Ser Cys Val
210 215 220
Glu Phe Glu Pro Glu Thr Val Pro Leu Leu Ser Thr Leu Arg Gly Lys
225 230 235 240
Pro Ile Thr Phe Leu Gly Leu Met Pro Pro Leu His Glu Gly Arg Arg
245 250 255
Glu Asp Gly Glu Asp Ala Thr Val Arg Trp Leu Asp Ala Gln Pro Ala
260 265 270
Lys Ser Val Val Tyr Val Ala Leu Gly Ser Glu Val Pro Leu Gly Val
275 280 285
Glu Lys Val His Glu Leu Ala Leu Gly Leu Glu Leu Ala Gly Thr Arg
290 295 300
Phe Leu Trp Ala Leu Arg Lys Pro Thr Gly Val Ser Asp Ala Asp Leu
305 310 315 320
Leu Pro Ala Gly Phe Glu Glu Arg Thr Arg Gly Arg Gly Val Val Ala
325 330 335
Thr Arg Trp Val Pro Gln Met Ser Ile Leu Ala His Ala Ala Val Gly
340 345 350
Ala Phe Leu Thr His Cys Gly Trp Asn Ser Thr Ile Glu Gly Leu Met
355 360 365
Phe Gly His Pro Leu Ile Met Leu Pro Ile Phe Gly Asp Gln Gly Pro
370 375 380
Asn Ala Arg Leu Ile Glu Ala Lys Asn Ala Gly Leu Gln Val Ala Arg
385 390 395 400
Asn Asp Gly Asp Gly Ser Phe Asp Arg Glu Gly Val Ala Ala Ala Ile
405 410 415
Arg Ala Val Ala Val Glu Glu Glu Ser Ser Lys Val Phe Gln Ala Lys
420 425 430
Ala Lys Lys Leu Gln Glu Ile Val Ala Asp Met Ala Cys His Glu Arg
435 440 445
Tyr Ile Asp Gly Phe Ile Gln Gln Leu Arg Ser Tyr Lys Asp
450 455 460
<210> 116
<211> 1389
<212> DNA
<213> 水稻
<400> 116
atggatagcg gttatagcag cagctatgca gcagcagccg gtatgcatgt tgttatttgt 60
ccgtggctgg catttggtca tctgctgccg tgtctggatc tggcacagcg tctggcaagc 120
cgtggtcatc gtgttagctt tgttagcaca ccgcgtaata ttagccgtct gcctccggtt 180
cgtccggcac tggcaccgct ggttgcattt gttgcactgc cgctgcctcg tgttgaaggt 240
ctgccggatg gtgcagaaag caccaatgat gttccgcatg atcgtccgga tatggttgaa 300
ctgcatcgtc gtgcatttga tggtctggca gcaccgttta gcgaatttct gggcaccgca 360
tgtgcagatt gggttattgt tgatgttttt catcattggg cagccgcagc agcactggaa 420
cataaagttc cgtgtgcaat gatgctgctg ggtagcgcac atatgattgc aagcattgca 480
gatcgtcgtc tggaacgtgc agaaaccgaa agtcctgcgg cagcaggtca gggtcgtcct 540
gcagccgcac cgacctttga agttgcacgt atgaaactga ttcgtaccaa aggtagcagc 600
ggtatgagcc tggcagaacg ttttagtctg accctgagcc gtagcagcct ggttgttggt 660
cgtagctgtg ttgaatttga accggaaacc gttccgctgc tgagcaccct gcgtggtaaa 720
ccgattacct ttctgggtct gatgcctccg ctgcatgaag gtcgtcgcga agatggtgaa 780
gatgcaaccg ttcgttggct ggatgcacag cctgcaaaaa gcgttgttta tgttgccctg 840
ggtagtgaag ttccgctggg tgttgaaaaa gtgcatgaac tggcactggg tttagaactg 900
gcaggcaccc gttttctgtg ggcactgcgt aaaccgaccg gtgttagtga tgccgatctg 960
cttccggcag gttttgaaga acgtacccgt ggtcgtggtg ttgttgcaac ccgttgggtt 1020
ccgcagatga gcattctggc acatgcagca gtgggtgcat ttctgaccca ttgtggttgg 1080
aatagcacca ttgaaggcct gatgtttggc catccgctga ttatgctgcc gatttttggt 1140
gatcagggtc cgaatgcacg tctgattgaa gcaaaaaatg caggtctgca ggttgcccgt 1200
aatgatggtg atggtagctt tgatcgtgaa ggtgttgcag cagccattcg tgcagttgca 1260
gttgaagaag aaagcagcaa agtttttcag gccaaagcca aaaaactgca agaaattgtt 1320
gcagatatgg cctgccatga acgttatatt gatggtttta ttcagcagct gcgtagctac 1380
aaagattaa 1389
<210> 117
<211> 487
<212> PRT
<213> 潘那利番茄(S. pennellii)
<400> 117
Met Gly Val Leu Thr Ile Glu Pro His Phe Val Leu Phe Pro Phe Met
1 5 10 15
Ala Gln Gly His Thr Ile Pro Met Ile Asp Ile Ala Arg Leu Leu Ala
20 25 30
Gln Arg Glu Val Ile Ile Thr Ile Val Thr Thr His Leu Asn Ala Asn
35 40 45
Arg Phe Lys Lys Val Ile Asp Arg Ala Ile Glu Ser Gly Leu Lys Ile
50 55 60
Gln Val Val His Leu Tyr Phe Pro Ser Leu Glu Ala Gly Leu Pro Glu
65 70 75 80
Gly Cys Glu Asn Phe Asp Met Leu Pro Ser Met Asp Leu Gly Leu Lys
85 90 95
Phe Phe Asp Ala Thr Lys Arg Leu Gln Pro Gln Val Glu Glu Met Leu
100 105 110
Gln Glu Met Lys Pro Ser Pro Ser Cys Ile Ile Ser Asp Met Cys Phe
115 120 125
Pro Trp Thr Thr Asn Val Ala Gln Lys Phe Asn Ile Pro Arg Ile Val
130 135 140
Phe His Gly Met Gly Cys Phe Ser Leu Leu Cys Leu His Asn Leu Lys
145 150 155 160
Asp Trp Glu Gly Leu Glu Lys Ile Glu Ser Asp Thr Glu Tyr Phe Gln
165 170 175
Val Pro Gly Leu Phe Asp Lys Ile Glu Leu Thr Lys Asn Gln Leu Gly
180 185 190
Asn Ala Ala Arg Pro Arg Asn Glu Glu Trp Arg Val Ile Ser Asp Gln
195 200 205
Met Lys Lys Ala Glu Glu Glu Ala Tyr Gly Met Val Val Asn Ser Phe
210 215 220
Glu Asp Leu Glu Lys Glu Tyr Ile Glu Gly Leu Met Asn Val Lys Asn
225 230 235 240
Arg Lys Ile Trp Thr Ile Gly Pro Val Ser Leu Cys Asn Lys Glu Lys
245 250 255
Gln Asp Lys Ala Glu Arg Gly Asn Lys Ala Ser Ile Asp Glu His Lys
260 265 270
Cys Leu Asn Trp Leu Asp Ser Arg Glu Gln Asn Ser Val Leu Phe Val
275 280 285
Cys Leu Gly Ser Leu Ser Arg Leu Ser Thr Ser Gln Met Val Glu Leu
290 295 300
Gly Leu Gly Leu Glu Ser Ser Arg Arg Pro Phe Ile Trp Val Val Arg
305 310 315 320
His Met Ser Asp Glu Phe Lys Asn Trp Leu Val Glu Glu Asp Phe Glu
325 330 335
Glu Arg Val Lys Gly Gln Gly Leu Leu Ile Arg Gly Trp Ala Pro Gln
340 345 350
Val Leu Ile Leu Ser His Pro Ser Ile Gly Ala Phe Leu Thr His Cys
355 360 365
Gly Trp Asn Ser Ser Leu Glu Gly Ile Thr Ala Gly Val Ala Met Ile
370 375 380
Thr Trp Pro Met Phe Ala Glu Gln Phe Cys Asn Glu Arg Leu Ile Val
385 390 395 400
Asp Val Leu Lys Thr Gly Val Arg Ser Gly Ile Glu Arg Gln Val Met
405 410 415
Phe Gly Glu Glu Glu Lys Leu Gly Thr Gln Val Ser Arg Asp Asp Ile
420 425 430
Lys Lys Val Ile Glu Gln Val Met Gly Glu Glu Met Arg Arg Lys Arg
435 440 445
Ala Lys Glu Leu Gly Glu Lys Ala Lys Arg Ala Met Glu Glu Glu Gly
450 455 460
Ser Ser His Phe Asn Leu Thr Gln Leu Ile Gln Asp Val Thr Glu Gln
465 470 475 480
Ala Lys Ile Leu Lys Pro Met
485
<210> 118
<211> 1464
<212> DNA
<213> 潘那利番茄
<400> 118
atgggtgttc tgaccattga accgcatttt gttctgtttc cgtttatggc acagggtcat 60
accattccga tgattgatat tgcacgtctg ctggcacagc gtgaagtgat tattaccatt 120
gttaccacac atctgaatgc caaccgtttc aaaaaagtta ttgatcgtgc aatcgagagc 180
ggtctgaaaa ttcaggttgt tcatctgtat tttccgagcc tggaagcagg tctgccggaa 240
ggttgtgaaa attttgatat gctgccgagc atggatctgg gtctgaaatt tttcgatgca 300
accaaacgtc tgcagccgca ggttgaagaa atgctgcaag aaatgaaacc gagtccgagc 360
tgtattatta gcgatatgtg ttttccgtgg accaccaatg ttgcacagaa atttaacatt 420
ccgcgtatcg tgtttcatgg tatgggttgt tttagcctgc tgtgtctgca taatctgaaa 480
gattgggaag gcctggaaaa aattgaaagc gataccgaat attttcaggt tccgggtctg 540
tttgataaaa tcgaactgac caaaaatcag ctgggtaatg cagcacgtcc gcgtaatgaa 600
gaatggcgtg tgattagcga tcagatgaaa aaagccgaag aagaggcata tggtatggtg 660
gttaatagct ttgaggatct ggaaaaagaa tacatcgaag gcctgatgaa tgtgaaaaac 720
cgtaaaattt ggaccattgg tccggttagc ctgtgcaata aagaaaaaca ggataaagcc 780
gaacgcggta ataaagcaag catcgatgaa cataaatgcc tgaattggct ggatagccgt 840
gaacagaata gcgttctgtt tgtttgtctg ggtagcctga gccgtctgag caccagccag 900
atggttgaat taggtctggg tttagaaagc agccgtcgtc cgtttatttg ggttgttcgt 960
catatgtccg atgagtttaa aaactggctg gtcgaagagg attttgaaga acgtgttaaa 1020
ggtcagggtc tgctgattcg tggttgggca ccgcaggttc tgattctgag ccatccgagc 1080
attggtgcat ttctgaccca ttgtggttgg aatagcagtc tggaaggtat taccgcaggc 1140
gttgcaatga ttacctggcc gatgtttgca gaacagtttt gtaatgaacg tctgattgtg 1200
gatgttctga aaaccggtgt tcgtagcggt attgaacgtc aggttatgtt tggtgaagaa 1260
gaaaaactgg gtacacaggt tagccgtgat gatatcaaaa aggtgattga acaggtgatg 1320
ggtgaagaga tgcgtcgtaa acgtgcaaaa gaactgggtg aaaaagcaaa acgtgccatg 1380
gaagaagaag gtagcagcca ttttaatctg acacagctga ttcaggatgt taccgaacag 1440
gcaaaaattc tgaaaccgat gtaa 1464
<210> 119
<211> 463
<212> PRT
<213> 水稻
<400> 119
Met Ala Ile Gly Ser Val Glu Ser Val Ala Val Val Ala Val Pro Phe
1 5 10 15
Pro Ala Gln Gly His Leu Asn Gln Leu Met His Leu Ser Leu Leu Leu
20 25 30
Ala Ser Arg Gly Leu Asp Val His Tyr Ala Ala Pro Pro Ala His Leu
35 40 45
Arg Gln Ala Arg Ser Arg Leu His Gly Trp Asp Pro Asp Ala Leu Arg
50 55 60
Ser Ile Arg Phe His Asp Leu Asp Val Pro Ala Tyr Glu Ser Pro Pro
65 70 75 80
Pro Asp Pro Thr Ala Pro Pro Phe Pro Ser His Met Met Pro Met Ile
85 90 95
Gln Ser Phe Ala Val Ala Ala Arg Ala Pro Phe Ala Ala Leu Leu Glu
100 105 110
Arg Ile Ser Ala Ser Tyr Ser Arg Val Val Val Val Tyr Asp Arg Leu
115 120 125
Asn Ser Phe Ala Ala Ala Gln Ala Ala Arg Leu Pro Asn Gly Glu Ala
130 135 140
Phe Gly Leu Gln Cys Val Ala Met Ser Tyr Asn Ile Gly Trp Leu Asp
145 150 155 160
Pro Glu Asn Arg Leu Val Arg Glu His Gly Leu Lys Phe His Pro Val
165 170 175
Glu Ala Cys Met Pro Lys Glu Phe Val Glu Phe Ile Ser Arg Glu Glu
180 185 190
Gln Asp Glu Glu Asn Ala Thr Ser Ser Gly Met Leu Met Asn Thr Ser
195 200 205
Arg Ala Ile Glu Ala Glu Phe Ile Asp Glu Ile Ala Ala His Pro Met
210 215 220
Phe Lys Glu Met Lys Leu Phe Ala Val Gly Pro Leu Asn Pro Leu Leu
225 230 235 240
Asp Ala Thr Ala Arg Thr Pro Gly Gln Thr Arg His Glu Cys Met Asp
245 250 255
Trp Leu Asp Lys Gln Pro Ala Ala Ser Val Leu Tyr Val Ser Phe Gly
260 265 270
Thr Thr Ser Ser Leu Arg Gly Asp Gln Val Ala Glu Leu Ala Ala Ala
275 280 285
Leu Lys Gly Ser Lys Gln Arg Phe Ile Trp Val Leu Arg Asp Ala Asp
290 295 300
Arg Ala Asp Ile Phe Ala Asp Ser Gly Glu Ser Arg His Ala Glu Leu
305 310 315 320
Leu Ser Arg Phe Thr Ala Glu Thr Glu Gly Val Gly Leu Val Ile Thr
325 330 335
Gly Trp Ala Pro Gln Leu Glu Ile Leu Ala His Gly Ala Thr Ala Ala
340 345 350
Phe Met Ser His Cys Gly Trp Asn Ser Thr Met Glu Ser Leu Ser His
355 360 365
Gly Lys Pro Ile Leu Ala Trp Pro Met His Ser Asp Gln Pro Trp Asp
370 375 380
Ala Glu Leu Val Cys Lys Tyr Leu Lys Ala Gly Leu Leu Val Arg Pro
385 390 395 400
Leu Glu Lys His Ser Glu Val Val Pro Ala Glu Ala Ile Gln Glu Val
405 410 415
Ile Glu Glu Ala Met Leu Pro Glu Lys Gly Met Ala Ile Arg Arg Arg
420 425 430
Ala Met Glu Leu Gly Glu Val Val Arg Ala Ser Val Ala Asp Gly Gly
435 440 445
Ser Ser Arg Lys Asp Leu Asp Asp Phe Val Gly Tyr Ile Thr Arg
450 455 460
<210> 120
<211> 1392
<212> DNA
<213> 水稻
<400> 120
atggcaattg gtagcgttga aagcgttgca gttgttgccg ttccgtttcc ggcacagggt 60
catctgaacc agctgatgca tctgagcctg ctgctggcaa gccgtggtct ggatgttcat 120
tatgcagcac cgcctgcaca tctgcgtcag gcacgtagcc gtctgcatgg ttgggatcct 180
gatgcactgc gtagcattcg ttttcatgat ctggatgtgc ctgcatatga aagtccgcct 240
ccggatccga ccgcaccgcc ttttccgagc catatgatgc cgatgattca gagctttgca 300
gttgcagcac gtgcaccgtt tgcagcactg ctggaacgta ttagcgcaag ctatagccgt 360
gttgttgttg tgtatgatcg tctgaatagc tttgccgcag cacaggcagc acgtctgccg 420
aatggtgaag catttggtct gcagtgtgtt gcaatgagct ataacattgg ttggctggat 480
ccggaaaatc gtctggttcg tgaacatggt ctgaaattcc atccggttga agcatgtatg 540
ccgaaagaat ttgttgaatt tatcagccgt gaagaacagg atgaagaaaa tgcaaccagc 600
agcggtatgc tgatgaatac cagccgtgca attgaagccg aatttattga tgaaattgca 660
gcgcacccga tgttcaaaga aatgaaactg tttgccgttg gtccgctgaa tcctctgctg 720
gatgcaaccg cacgtacacc gggtcagacc cgtcatgaat gtatggattg gctggacaaa 780
cagcctgcag caagcgttct gtatgttagc tttggcacca ccagtagcct gcgtggtgat 840
caggttgcag aactggcagc agcactgaaa ggtagcaaac agcgttttat ttgggttctg 900
cgtgatgcag atcgtgcaga tatttttgca gatagcggtg aaagccgtca tgccgaactg 960
ctgagccgtt ttaccgcaga aaccgaaggt gttggtctgg ttattaccgg ttgggcaccg 1020
cagctggaaa ttctggcaca tggtgccacc gcagcattta tgagccattg tggttggaat 1080
agcaccatgg aaagcctgag ccatggtaaa ccgattctgg catggccgat gcatagcgat 1140
cagccttggg atgctgaact ggtttgtaaa tatctgaaag caggtctgct ggttcgtccg 1200
ctggaaaaac atagcgaagt tgttccggca gaagcaattc aagaagttat tgaagaagca 1260
atgctgccgg aaaaaggtat ggcaattcgt cgtcgtgcaa tggaactggg tgaagttgtg 1320
cgtgcaagcg ttgccgatgg tggtagcagc cgtaaagatc tggacgattt tgttggttat 1380
atcacccgct aa 1392
<210> 121
<211> 456
<212> PRT
<213> 拟南芥
<400> 121
Met Gly Ser Ser Glu Gly Gln Glu Thr His Val Leu Met Val Thr Leu
1 5 10 15
Pro Phe Gln Gly His Ile Asn Pro Met Leu Lys Leu Ala Lys His Leu
20 25 30
Ser Leu Ser Ser Lys Asn Leu His Ile Asn Leu Ala Thr Ile Glu Ser
35 40 45
Ala Arg Asp Leu Leu Ser Thr Val Glu Lys Pro Arg Tyr Pro Val Asp
50 55 60
Leu Val Phe Phe Ser Asp Gly Leu Pro Lys Glu Asp Pro Lys Ala Pro
65 70 75 80
Glu Thr Leu Leu Lys Ser Leu Asn Lys Val Gly Ala Met Asn Leu Ser
85 90 95
Lys Ile Ile Glu Glu Lys Arg Tyr Ser Cys Ile Ile Ser Ser Pro Phe
100 105 110
Thr Pro Trp Val Pro Ala Val Ala Ala Ser His Asn Ile Ser Cys Ala
115 120 125
Ile Leu Trp Ile Gln Ala Cys Gly Ala Tyr Ser Val Tyr Tyr Arg Tyr
130 135 140
Tyr Met Lys Thr Asn Ser Phe Pro Asp Leu Glu Asp Leu Asn Gln Thr
145 150 155 160
Val Glu Leu Pro Ala Leu Pro Leu Leu Glu Val Arg Asp Leu Pro Ser
165 170 175
Phe Met Leu Pro Ser Gly Gly Ala His Phe Tyr Asn Leu Met Ala Glu
180 185 190
Phe Ala Asp Cys Leu Arg Tyr Val Lys Trp Val Leu Val Asn Ser Phe
195 200 205
Tyr Glu Leu Glu Ser Glu Ile Ile Glu Ser Met Ala Asp Leu Lys Pro
210 215 220
Val Ile Pro Ile Gly Pro Leu Val Ser Pro Phe Leu Leu Gly Asp Gly
225 230 235 240
Glu Glu Glu Thr Leu Asp Gly Lys Asn Leu Asp Phe Cys Lys Ser Asp
245 250 255
Asp Cys Cys Met Glu Trp Leu Asp Lys Gln Ala Arg Ser Ser Val Val
260 265 270
Tyr Ile Ser Phe Gly Ser Met Leu Glu Thr Leu Glu Asn Gln Val Glu
275 280 285
Thr Ile Ala Lys Ala Leu Lys Asn Arg Gly Leu Pro Phe Leu Trp Val
290 295 300
Ile Arg Pro Lys Glu Lys Ala Gln Asn Val Ala Val Leu Gln Glu Met
305 310 315 320
Val Lys Glu Gly Gln Gly Val Val Leu Glu Trp Ser Pro Gln Glu Lys
325 330 335
Ile Leu Ser His Glu Ala Ile Ser Cys Phe Val Thr His Cys Gly Trp
340 345 350
Asn Ser Thr Met Glu Thr Val Val Ala Gly Val Pro Val Val Ala Tyr
355 360 365
Pro Ser Trp Thr Asp Gln Pro Ile Asp Ala Arg Leu Leu Val Asp Val
370 375 380
Phe Gly Ile Gly Val Arg Met Arg Asn Asp Ser Val Asp Gly Glu Leu
385 390 395 400
Lys Val Glu Glu Val Glu Arg Cys Ile Glu Ala Val Thr Glu Gly Pro
405 410 415
Ala Ala Val Asp Ile Arg Arg Arg Ala Ala Glu Leu Lys Arg Val Ala
420 425 430
Arg Leu Ala Leu Ala Pro Gly Gly Ser Ser Thr Arg Asn Leu Asp Leu
435 440 445
Phe Ile Ser Asp Ile Thr Ile Ala
450 455
<210> 122
<211> 1371
<212> DNA
<213> 拟南芥
<400> 122
atgggtagca gcgaaggtca agaaacccat gttctgatgg ttaccctgcc gtttcagggt 60
catattaatc cgatgctgaa actggcaaaa catctgagcc tgagcagcaa aaatctgcat 120
attaacctgg caaccattga aagcgcacgt gatctgctga gcaccgttga aaaaccgcgt 180
tatccggttg atctggtgtt ttttagtgat ggtctgccga aagaagatcc gaaagcaccg 240
gaaacactgc tgaaaagcct gaataaagtt ggtgcaatga acctgagcaa aatcatcgaa 300
gaaaaacgct atagctgcat tattagcagc ccgtttacac cgtgggttcc agcagttgca 360
gcaagccata acattagctg tgcaattctg tggattcagg catgtggtgc atatagcgtg 420
tattatcgct attatatgaa aaccaacagc ttcccggatc tggaagatct gaatcagacc 480
gttgaactgc ctgcactgcc gctgctggaa gttcgcgatc tgccgagctt tatgctgccg 540
agcggtggtg cacatttcta taatctgatg gcagaatttg cagattgcct gcgttatgtt 600
aaatgggtgt tagtgaacag cttctatgaa ctggaaagcg aaattattga aagcatggca 660
gatctgaaac cggttattcc gattggtccg ctggttagcc cgtttctgtt aggtgatggt 720
gaagaagaaa ccctggacgg taaaaatctg gatttttgta aatccgatga ttgctgcatg 780
gaatggctgg ataaacaggc acgtagcagc gttgtgtata ttagctttgg tagcatgctg 840
gaaacgctgg aaaatcaggt tgaaaccatt gcaaaagccc tgaaaaatcg cggtctgcct 900
tttctgtggg ttattcgtcc gaaagaaaaa gcacagaatg ttgcagttct gcaagagatg 960
gttaaagaag gtcagggcgt tgttctggaa tggtcaccgc aagaaaaaat tctgagccat 1020
gaagcgatta gctgctttgt tacccattgt ggttggaata gcaccatgga aaccgttgtt 1080
gccggtgttc cggttgttgc atatccgagc tggaccgatc agccgattga tgcacgtctg 1140
ctggttgatg tttttggtat tggtgttcgt atgcgtaatg atagcgtgga tggtgaactg 1200
aaagttgaag aagttgaacg ttgtattgaa gccgttaccg aaggtccggc agcagttgat 1260
attcgtcgtc gtgcagcaga actgaaacgt gttgcccgtc tggcactggc acctggtggt 1320
agcagcaccc gtaatctgga cctgtttatt agcgatatta ccattgccta a 1371
<210> 123
<211> 483
<212> PRT
<213> 甜叶菊
<400> 123
Met Asp Gln Met Ala Lys Ile Asp Glu Lys Lys Pro His Val Val Phe
1 5 10 15
Ile Pro Phe Pro Ala Gln Ser His Ile Lys Cys Met Leu Lys Leu Ala
20 25 30
Arg Ile Leu His Gln Lys Gly Leu Tyr Ile Thr Phe Ile Asn Thr Asp
35 40 45
Thr Asn His Glu Arg Leu Val Ala Ser Gly Gly Thr Gln Trp Leu Glu
50 55 60
Asn Ala Pro Gly Phe Trp Phe Lys Thr Val Pro Asp Gly Phe Gly Ser
65 70 75 80
Ala Lys Asp Asp Gly Val Lys Pro Thr Asp Ala Leu Arg Glu Leu Met
85 90 95
Asp Tyr Leu Lys Thr Asn Phe Phe Asp Leu Phe Leu Asp Leu Val Leu
100 105 110
Lys Leu Glu Val Pro Ala Thr Cys Ile Ile Cys Asp Gly Cys Met Thr
115 120 125
Phe Ala Asn Thr Ile Arg Ala Ala Glu Lys Leu Asn Ile Pro Val Ile
130 135 140
Leu Phe Trp Thr Met Ala Ala Cys Gly Phe Met Ala Phe Tyr Gln Ala
145 150 155 160
Lys Val Leu Lys Glu Lys Glu Ile Val Pro Val Lys Asp Glu Thr Tyr
165 170 175
Leu Thr Asn Gly Tyr Leu Asp Met Glu Ile Asp Trp Ile Pro Gly Met
180 185 190
Lys Arg Ile Arg Leu Arg Asp Leu Pro Glu Phe Ile Leu Ala Thr Lys
195 200 205
Gln Asn Tyr Phe Ala Phe Glu Phe Leu Phe Glu Thr Ala Gln Leu Ala
210 215 220
Asp Lys Val Ser His Met Ile Ile His Thr Phe Glu Glu Leu Glu Ala
225 230 235 240
Ser Leu Val Ser Glu Ile Lys Ser Ile Phe Pro Asn Val Tyr Thr Ile
245 250 255
Gly Pro Leu Gln Leu Leu Leu Asn Lys Ile Thr Gln Lys Glu Thr Asn
260 265 270
Asn Asp Ser Tyr Ser Leu Trp Lys Glu Glu Pro Glu Cys Val Glu Trp
275 280 285
Leu Asn Ser Lys Glu Pro Asn Ser Val Val Tyr Val Asn Phe Gly Ser
290 295 300
Leu Ala Val Met Ser Leu Gln Asp Leu Val Glu Phe Gly Trp Gly Leu
305 310 315 320
Val Asn Ser Asn His Tyr Phe Leu Trp Ile Ile Arg Ala Asn Leu Ile
325 330 335
Asp Gly Lys Pro Ala Val Met Pro Gln Glu Leu Lys Glu Ala Met Asn
340 345 350
Glu Lys Gly Phe Val Gly Ser Trp Cys Ser Gln Glu Glu Val Leu Asn
355 360 365
His Pro Ala Val Gly Gly Phe Leu Thr His Cys Gly Trp Gly Ser Ile
370 375 380
Ile Glu Ser Leu Ser Ala Gly Val Pro Met Leu Gly Trp Pro Ser Ile
385 390 395 400
Gly Asp Gln Arg Ala Asn Cys Arg Gln Met Cys Lys Glu Trp Glu Val
405 410 415
Gly Met Glu Ile Gly Lys Asn Val Lys Arg Asp Glu Val Glu Lys Leu
420 425 430
Val Arg Met Leu Met Glu Gly Leu Glu Gly Glu Arg Met Arg Lys Lys
435 440 445
Ala Leu Glu Trp Lys Lys Ser Ala Thr Leu Ala Thr Cys Cys Asn Gly
450 455 460
Ser Ser Ser Leu Asp Val Glu Lys Leu Ala Asn Glu Ile Lys Lys Leu
465 470 475 480
Ser Arg Asn
<210> 124
<211> 1452
<212> DNA
<213> 甜叶菊
<400> 124
atggatcaga tggccaaaat cgatgaaaaa aaaccgcatg tggtgtttat tccgtttccg 60
gcacagagcc atatcaaatg tatgctgaaa ctggcacgta tcctgcatca gaaaggtctg 120
tatattacct tcattaacac cgataccaat catgaacgtc tggttgcaag cggtggcacc 180
cagtggctgg aaaatgcacc tggtttttgg tttaaaaccg ttccggatgg ttttggtagc 240
gcaaaagatg atggtgttaa accgaccgat gcactgcgtg aactgatgga ttatctgaaa 300
accaactttt tcgacctgtt tctggatctg gtgctgaaat tagaagttcc ggcaacctgt 360
attatttgtg atggttgtat gacctttgcc aataccattc gtgcagcaga aaaactgaat 420
attccggtga ttctgttttg gaccatggca gcctgtggtt ttatggcatt ttatcaggca 480
aaagtgctga aagaaaaaga aatcgttccg gtgaaagatg aaacctatct gaccaatggt 540
tatctggata tggaaatcga ttggattccg ggtatgaaac gtattcgtct gcgtgatctg 600
ccggaattta ttctggcaac caaacagaac tatttcgcct ttgaatttct gttcgaaacc 660
gcacagctgg cagataaagt tagccatatg attatccaca ccttcgaaga actggaagca 720
agcctggtta gcgaaatcaa aagcattttt ccgaacgtgt atacaattgg tccgctgcag 780
ctgctgctga acaaaattac ccagaaagaa accaacaacg atagctatag cctgtggaaa 840
gaagaaccgg aatgtgttga atggctgaat agcaaagaac cgaatagcgt tgtgtatgtg 900
aattttggta gtctggcagt tatgagcctg caggatctgg ttgaatttgg ttggggttta 960
gttaacagca accactattt tctgtggatt attcgtgcca atctgattga tggtaaaccg 1020
gcagtgatgc cgcaagaact gaaagaagca atgaacgaaa aaggttttgt tggtagctgg 1080
tgtagccaag aagaagttct gaatcatccg gcagttggtg gttttctgac ccattgcggt 1140
tggggtagca ttattgaaag cctgagtgcc ggtgttccga tgttaggttg gccgagcatt 1200
ggtgatcagc gtgcaaattg tcgtcagatg tgtaaagaat gggaagttgg tatggaaatt 1260
ggcaaaaacg tgaaacgtga tgaggttgaa aaactggttc gtatgctgat ggaaggtctg 1320
gaaggtgaac gtatgcgtaa aaaagcactg gaatggaaaa aaagcgcaac cctggccacc 1380
tgttgtaatg gtagcagcag cctggatgtt gagaaactgg ccaatgaaat taagaaactg 1440
agccgcaact aa 1452
<210> 125
<211> 498
<212> PRT
<213> P. abies
<400> 125
Met Asn Gly Asn Glu Gln His Ala Leu His Ala Val Ile Val Pro Phe
1 5 10 15
Pro Ala Gln Gly His Val Asn Ala Leu Met Asn Leu Ala Gln Leu Leu
20 25 30
Ala Ile Arg Gly Val Phe Val Thr Phe Val Asn Thr Asp Trp Ile His
35 40 45
Lys Arg Thr Val Glu Ala Ser Lys Lys Ser Lys Ser Gly Val Leu Asn
50 55 60
Asp Asn Pro Glu Phe Glu Gln Gln Gly Arg Arg Ile Arg Phe Leu Ser
65 70 75 80
Ile Pro Asp Gly Leu Pro Pro Gly Asp Gly Arg Thr Ser Asn Leu Gly
85 90 95
Glu Leu Phe Val Ala Leu Gln Lys Leu Gly Pro Val Leu Glu Asp Leu
100 105 110
Leu Arg Thr Ala Asp Glu Lys Ser Pro Ser Phe Pro Pro Ile Thr Phe
115 120 125
Ile Val Thr Asp Ala Phe Met Ser Cys Thr Glu Gln Val Ala Ser Ser
130 135 140
Met Lys Val Pro Arg Val Ile Phe Trp Pro Val Cys Ala Ala Ile Ser
145 150 155 160
Ile Ser Gln Tyr Tyr Ala Asp Leu Leu Ile Ser Glu Gly Tyr Ile Pro
165 170 175
Val Asn Leu Ser Gln Ala Lys Asn Pro Glu Lys Leu Ile Thr Cys Leu
180 185 190
Pro Gly Asn Ile Pro Pro Leu Lys Pro Thr Asp Leu Val Ser Phe Tyr
195 200 205
Arg Ala Gln Asp Pro Thr Asp Ile Leu Phe Asn Ala Phe Leu His Glu
210 215 220
Ser Arg Lys Gln Ser Lys Gly Asp Tyr Val Leu Val Asn Thr Phe Glu
225 230 235 240
Glu Leu Glu Gly Arg Asp Ala Val Thr Ala Leu Ser Leu Asp Gly Cys
245 250 255
Pro Ala Leu Ala Ile Gly Pro Leu Phe Leu Pro Asn Phe Leu Glu Gly
260 265 270
Arg Asp Ser Cys Ser Ser Leu Trp Glu Glu Glu Lys Ser Cys Leu Thr
275 280 285
Trp Leu Asp Met His Gln Pro Gly Ser Val Ile Tyr Val Ser Phe Gly
290 295 300
Ser Ile Ala Val Lys Ser Glu Gln Gln Leu Glu Gln Leu Ala Leu Gly
305 310 315 320
Leu Glu Gly Ser Gly Gln Pro Phe Leu Trp Val Leu Arg Leu Asp Ile
325 330 335
Ala Glu Gly Gln Ala Ala Val Leu Pro Asp Gly Phe Glu Ala Arg Thr
340 345 350
Lys Asp Arg Ala Leu Phe Val Arg Trp Ala Pro Gln Trp Asn Val Leu
355 360 365
Ala His Pro Ser Val Gly Leu Phe Leu Thr His Cys Gly Trp Asn Ser
370 375 380
Thr Leu Glu Ser Met Ser Met Gly Val Pro Val Val Gly Phe Pro Tyr
385 390 395 400
Phe Gly Asp Gln Phe Leu Asn Cys Arg Phe Ala Lys Asp Val Trp Arg
405 410 415
Ile Gly Leu Asp Phe Lys Asp Val Asp Leu Asp Asp Arg Lys Val Val
420 425 430
Met Lys Glu Glu Val Glu Asp Val Val Arg Arg Met Met Arg Thr Pro
435 440 445
Glu Gly Lys Lys Leu Arg Asp Asn Val Leu Arg Leu Lys Glu Ser Ala
450 455 460
Ala Lys Ala Val Leu Pro Gly Gly Ser Ser Phe Leu Asn Leu Asn Thr
465 470 475 480
Phe Val Lys Asp Met Thr Thr Gly Lys Gly Phe Gln Ser Lys Asn Glu
485 490 495
Thr Met
<210> 126
<211> 1497
<212> DNA
<213> P. abies
<400> 126
atgaatggca atgaacagca tgccctgcat gccgttattg ttccgtttcc ggcacagggt 60
catgttaatg cactgatgaa tctggcacag ctgctggcaa ttcgtggtgt ttttgttacc 120
tttgttaaca ccgattggat ccataaacgt accgttgaag caagcaaaaa aagcaaaagc 180
ggtgtgctga atgataaccc ggaatttgaa cagcagggtc gtcgtattcg ttttctgagc 240
attccggatg gtctgcctcc aggtgatggt cgtaccagca atctgggtga actgtttgtt 300
gcactgcaga aactgggtcc tgttctggaa gatctgctgc gtaccgcaga tgaaaaaagc 360
ccgagctttc cgcctattac ctttattgtt accgatgcct ttatgagctg taccgaacag 420
gttgcaagca gcatgaaagt tccgcgtgtg attttttggc ctgtttgtgc agcaattagc 480
atcagccagt attatgccga tctgctgatt agcgaaggtt atattccggt taatctgagc 540
caggcgaaaa atccggaaaa actgattacc tgtctgcctg gtaatattcc gcctctgaaa 600
ccgaccgatc tggttagctt ttatcgtgca caggatccga ccgatattct gtttaatgca 660
tttctgcatg aaagccgcaa acagagcaaa ggtgattatg ttctggtgaa cacctttgaa 720
gaactggaag gtcgtgatgc agttaccgca ctgagcctgg atggttgtcc ggcactggca 780
attggtccgc tgtttctgcc gaattttctg gaaggacgcg atagctgtag cagcctgtgg 840
gaagaagaaa aaagctgtct gacctggctg gatatgcatc agcctggtag cgttatttat 900
gttagctttg gtagcattgc cgtgaaaagc gaacagcagc tggaacagct ggcactgggt 960
ttagaaggta gcggtcagcc gtttctgtgg gttctgcgtc tggatattgc agaaggtcag 1020
gcagcagttc tgccggatgg ttttgaagca cgtaccaaag atcgtgccct gtttgttcgt 1080
tgggcaccgc agtggaatgt tctggcacat ccgagcgttg gtctgtttct gacccattgt 1140
ggttggaata gcaccctgga aagcatgagc atgggtgttc cggttgttgg ttttccgtat 1200
tttggtgatc agtttctgaa ttgccgtttc gcaaaagatg tttggcgtat tggtctggat 1260
ttcaaagatg ttgatctgga tgatcgtaaa gtggtgatga aagaagaagt tgaggacgtt 1320
gttcgtcgta tgatgcgtac accggaaggt aaaaaactgc gtgataatgt gctgcgtctg 1380
aaagaaagcg cagcaaaagc cgttctgcca ggtggtagca gctttctgaa tctgaatacc 1440
tttgtgaaag atatgaccac cggtaaaggt ttccagagca aaaatgaaac catgtaa 1497
<210> 127
<211> 487
<212> PRT
<213> C. roseus
<400> 127
Met Val Asn Gln Leu His Ile Phe Asn Phe Pro Phe Met Ala Gln Gly
1 5 10 15
His Met Leu Pro Ala Leu Asp Met Ala Asn Leu Phe Thr Ser Arg Gly
20 25 30
Val Lys Val Thr Leu Ile Thr Thr His Gln His Val Pro Met Phe Thr
35 40 45
Lys Ser Ile Glu Arg Ser Arg Asn Ser Gly Phe Asp Ile Ser Ile Gln
50 55 60
Ser Ile Lys Phe Pro Ala Ser Glu Val Gly Leu Pro Glu Gly Ile Glu
65 70 75 80
Ser Leu Asp Gln Val Ser Gly Asp Asp Glu Met Leu Pro Lys Phe Met
85 90 95
Arg Gly Val Asn Leu Leu Gln Gln Pro Leu Glu Gln Leu Leu Gln Glu
100 105 110
Ser Arg Pro His Cys Leu Leu Ser Asp Met Phe Phe Pro Trp Thr Thr
115 120 125
Glu Ser Ala Ala Lys Phe Gly Ile Pro Arg Leu Leu Phe His Gly Ser
130 135 140
Cys Ser Phe Ala Leu Ser Ala Ala Glu Ser Val Arg Arg Asn Lys Pro
145 150 155 160
Phe Glu Asn Val Ser Thr Asp Thr Glu Glu Phe Val Val Pro Asp Leu
165 170 175
Pro His Gln Ile Lys Leu Thr Arg Thr Gln Ile Ser Thr Tyr Glu Arg
180 185 190
Glu Asn Ile Glu Ser Asp Phe Thr Lys Met Leu Lys Lys Val Arg Asp
195 200 205
Ser Glu Ser Thr Ser Tyr Gly Val Val Val Asn Ser Phe Tyr Glu Leu
210 215 220
Glu Pro Asp Tyr Ala Asp Tyr Tyr Ile Asn Val Leu Gly Arg Lys Ala
225 230 235 240
Trp His Ile Gly Pro Phe Leu Leu Cys Asn Lys Leu Gln Ala Glu Asp
245 250 255
Lys Ala Gln Arg Gly Lys Lys Ser Ala Ile Asp Ala Asp Glu Cys Leu
260 265 270
Asn Trp Leu Asp Ser Lys Gln Pro Asn Ser Val Ile Tyr Leu Cys Phe
275 280 285
Gly Ser Met Ala Asn Leu Asn Ser Ala Gln Leu His Glu Ile Ala Thr
290 295 300
Ala Leu Glu Ser Ser Gly Gln Asn Phe Ile Trp Val Val Arg Lys Cys
305 310 315 320
Val Asp Glu Glu Asn Ser Ser Lys Trp Phe Pro Glu Gly Phe Glu Glu
325 330 335
Arg Thr Lys Glu Lys Gly Leu Ile Ile Lys Gly Trp Ala Pro Gln Thr
340 345 350
Leu Ile Leu Glu His Glu Ser Val Gly Ala Phe Val Thr His Cys Gly
355 360 365
Trp Asn Ser Thr Leu Glu Gly Ile Cys Ala Gly Val Pro Leu Val Thr
370 375 380
Trp Pro Phe Phe Ala Glu Gln Phe Phe Asn Glu Lys Leu Ile Thr Glu
385 390 395 400
Val Leu Lys Thr Gly Tyr Gly Val Gly Ala Arg Gln Trp Ser Arg Val
405 410 415
Ser Thr Glu Ile Ile Lys Gly Glu Ala Ile Ala Asn Ala Ile Asn Arg
420 425 430
Val Met Val Gly Asp Glu Ala Val Glu Met Arg Asn Arg Ala Lys Asp
435 440 445
Leu Lys Glu Lys Ala Arg Lys Ala Leu Glu Glu Asp Gly Ser Ser Tyr
450 455 460
Arg Asp Leu Thr Ala Leu Ile Glu Glu Leu Gly Ala Tyr Arg Ser Gln
465 470 475 480
Val Glu Arg Lys Gln Gln Asp
485
<210> 128
<211> 1464
<212> DNA
<213> 长春花(C. roseus)
<400> 128
atggtgaacc agctgcacat ttttaacttt ccgtttatgg cacagggtca tatgctgcct 60
gcactggata tggcaaacct gtttaccagc cgtggtgtta aagttaccct gattaccaca 120
catcagcatg ttccgatgtt taccaaaagc attgaacgta gccgtaatag cggttttgat 180
attagcattc agagcatcaa atttccggca agcgaagttg gtctgccgga aggtattgaa 240
agcctggatc aggttagcgg tgatgatgaa atgctgccga aatttatgcg tggtgtgaat 300
ctgctgcaac agccgctgga acagctgctg caagaaagcc gtccgcattg tctgctgagc 360
gatatgtttt ttccgtggac caccgaaagc gcagcaaaat ttggtattcc gcgtctgctg 420
tttcatggta gctgtagctt tgcactgagc gcagcagaaa gcgttcgtcg taataaaccg 480
tttgaaaatg ttagcaccga taccgaagaa tttgttgttc cggatctgcc gcatcagatt 540
aaactgaccc gtacacagat tagcacctat gaacgtgaaa acatcgaaag cgatttcacc 600
aagatgctga aaaaagttcg tgatagcgaa agcaccagct atggtgttgt tgtgaatagc 660
ttttatgaac tggaaccgga ttatgccgat tactatatta acgttctggg tcgtaaagcc 720
tggcatattg gtccgtttct gctgtgtaat aaactgcagg ccgaagataa agcacagcgt 780
ggtaaaaaaa gcgcaattga tgcagatgaa tgtctgaatt ggctggatag caaacagccg 840
aatagcgtta tttatctgtg ttttggtagc atggccaatc tgaatagcgc acagctgcat 900
gaaattgcaa ccgcactgga aagcagcggt cagaacttta tttgggttgt tcgtaaatgc 960
gtggatgaag aaaatagcag caaatggttt ccggaaggct ttgaagaacg taccaaagaa 1020
aaaggcctga ttatcaaagg ttgggcaccg cagacactga ttctggaaca tgaaagcgtt 1080
ggtgcatttg ttacccattg tggttggaat agcaccctgg aaggcatttg tgccggtgtt 1140
ccgctggtta cctggccgtt ttttgcagaa cagtttttta acgagaaact gatcacggaa 1200
gttctgaaaa ccggttatgg tgtgggtgca cgtcagtggt cacgtgtgag caccgaaatc 1260
attaaaggtg aagcaattgc caatgccatt aatcgtgtta tggttggtga tgaagcagtg 1320
gaaatgcgta atcgtgcaaa agatctgaaa gagaaagcac gtaaagcact ggaagaagat 1380
ggtagcagct atcgtgatct gaccgcactg attgaagaac tgggtgcata tcgtagccag 1440
gttgaacgta aacagcagga ttaa 1464
<210> 129
<211> 481
<212> PRT
<213> 拟南芥
<400> 129
Met Ser Ser Asp Pro His Arg Lys Leu His Val Val Phe Phe Pro Phe
1 5 10 15
Met Ala Tyr Gly His Met Ile Pro Thr Leu Asp Met Ala Lys Leu Phe
20 25 30
Ser Ser Arg Gly Ala Lys Ser Thr Ile Leu Thr Thr Pro Leu Asn Ser
35 40 45
Lys Ile Phe Gln Lys Pro Ile Glu Arg Phe Lys Asn Leu Asn Pro Ser
50 55 60
Phe Glu Ile Asp Ile Gln Ile Phe Asp Phe Pro Cys Val Asp Leu Gly
65 70 75 80
Leu Pro Glu Gly Cys Glu Asn Val Asp Phe Phe Thr Ser Asn Asn Asn
85 90 95
Asp Asp Arg Gln Tyr Leu Thr Leu Lys Phe Phe Lys Ser Thr Arg Phe
100 105 110
Phe Lys Asp Gln Leu Glu Lys Leu Leu Glu Thr Thr Arg Pro Asp Cys
115 120 125
Leu Ile Ala Asp Met Phe Phe Pro Trp Ala Thr Glu Ala Ala Glu Lys
130 135 140
Phe Asn Val Pro Arg Leu Val Phe His Gly Thr Gly Tyr Phe Ser Leu
145 150 155 160
Cys Ser Glu Tyr Cys Ile Arg Val His Asn Pro Gln Asn Ile Val Ala
165 170 175
Ser Arg Tyr Glu Pro Phe Val Ile Pro Asp Leu Pro Gly Asn Ile Val
180 185 190
Ile Thr Gln Glu Gln Ile Ala Asp Arg Asp Glu Glu Ser Glu Met Gly
195 200 205
Lys Phe Met Ile Glu Val Lys Glu Ser Asp Val Lys Ser Ser Gly Val
210 215 220
Ile Val Asn Ser Phe Tyr Glu Leu Glu Pro Asp Tyr Ala Asp Phe Tyr
225 230 235 240
Lys Ser Val Val Leu Lys Arg Ala Trp His Ile Gly Pro Leu Ser Val
245 250 255
Tyr Asn Arg Gly Phe Glu Glu Lys Ala Glu Arg Gly Lys Lys Ala Ser
260 265 270
Ile Asn Glu Val Glu Cys Leu Lys Trp Leu Asp Ser Lys Lys Pro Asp
275 280 285
Ser Val Ile Tyr Ile Ser Phe Gly Ser Val Ala Cys Phe Lys Asn Glu
290 295 300
Gln Leu Phe Glu Ile Ala Ala Gly Leu Glu Thr Ser Gly Ala Asn Phe
305 310 315 320
Ile Trp Val Val Arg Lys Asn Ile Gly Ile Glu Lys Glu Glu Trp Leu
325 330 335
Pro Glu Gly Phe Glu Glu Arg Val Lys Gly Lys Gly Met Ile Ile Arg
340 345 350
Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Gln Ala Thr Cys Gly
355 360 365
Phe Val Thr His Cys Gly Trp Asn Ser Leu Leu Glu Gly Val Ala Ala
370 375 380
Gly Leu Pro Met Val Thr Trp Pro Val Ala Ala Glu Gln Phe Tyr Asn
385 390 395 400
Glu Lys Leu Val Thr Gln Val Leu Arg Thr Gly Val Ser Val Gly Ala
405 410 415
Lys Lys Asn Val Arg Thr Thr Gly Asp Phe Ile Ser Arg Glu Lys Val
420 425 430
Val Lys Ala Val Arg Glu Val Leu Val Gly Glu Glu Ala Asp Glu Arg
435 440 445
Arg Glu Arg Ala Lys Lys Leu Ala Glu Met Ala Lys Ala Ala Val Glu
450 455 460
Gly Gly Ser Ser Phe Asn Asp Leu Asn Ser Phe Ile Glu Glu Phe Thr
465 470 475 480
Ser
<210> 130
<211> 1446
<212> DNA
<213> 拟南芥
<400> 130
atgagcagcg atccgcatcg taaactgcat gttgtttttt ttccgtttat ggcctatggt 60
catatgattc cgacactgga tatggcaaaa ctgtttagca gccgtggtgc aaaaagcacc 120
attctgacca caccgctgaa tagcaaaatc tttcagaaac cgattgagcg cttcaaaaat 180
ctgaatccga gctttgaaat cgacatccag atctttgatt ttccgtgtgt tgatctgggt 240
ctgccggaag gttgtgaaaa tgttgatttt ttcaccagca acaacaacga tgatcgtcag 300
tatctgaccc tgaaattttt caaaagcacc cgctttttca aagatcagct ggaaaaactg 360
ctggaaacca cacgtccgga ttgtctgatt gcagatatgt tttttccttg ggcaaccgaa 420
gcagccgaaa aattcaatgt tccgcgtctg gtttttcatg gcaccggtta ttttagcctg 480
tgtagcgaat attgcattcg tgttcataat ccgcagaata ttgttgccag ccgttatgaa 540
ccgtttgtga ttccggatct gcctggtaat attgttatta cccaagagca gattgccgat 600
cgtgatgaag aaagcgaaat gggcaaattt atgatcgaag ttaaagagag cgacgtcaaa 660
agcagcggtg ttattgttaa cagcttttat gaactggaac cggattatgc cgatttctat 720
aaaagcgttg ttctgaaacg tgcctggcat attggtccgc tgagcgttta taatcgtggc 780
tttgaagaaa aagccgagcg tggtaaaaaa gccagcatta atgaagttga atgcctgaaa 840
tggctggaca gcaaaaaacc ggatagcgtt atctatatta gctttggtag cgttgcctgc 900
tttaaaaacg agcagctgtt tgaaattgca gcaggtctgg aaacctcagg tgcaaacttt 960
atttgggttg tgcgtaaaaa catcggcatc gaaaaagaag aatggctgcc tgaaggtttt 1020
gaggaacgtg ttaaaggtaa aggcatgatt attcgtggtt gggcaccgca ggttctgatt 1080
ctggatcatc aggcaacctg tggttttgtt acccattgtg gttggaatag cctgctggaa 1140
ggtgtggcag ccggtctgcc gatggttacc tggcctgttg cagcagaaca gttttataac 1200
gaaaaactgg ttacccaggt tctgcgtacc ggtgttagcg ttggtgccaa aaaaaacgtt 1260
cgtaccaccg gtgatttcat cagccgtgaa aaagttgtta aagccgttcg tgaagttctg 1320
gttggtgaag aggcagatga acgtcgtgaa cgtgcaaaaa aactggcaga aatggcaaaa 1380
gccgcagttg aaggtggtag cagctttaat gatctgaaca gctttatcga agagtttacc 1440
agctaa 1446
<210> 131
<211> 474
<212> PRT
<213> 人工
<220>
<223> 人工
<400> 131
Met Gly Lys Gln Glu Asp Ala Glu Leu Val Ile Ile Pro Phe Pro Phe
1 5 10 15
Ser Gly His Ile Leu Ala Thr Ile Glu Leu Ala Lys Arg Leu Ile Ser
20 25 30
Gln Asp Asn Pro Arg Ile His Thr Ile Thr Ile Leu Tyr Trp Gly Leu
35 40 45
Pro Phe Ile Pro Gln Ala Asp Thr Ile Ala Phe Leu Arg Ser Leu Val
50 55 60
Lys Asn Glu Pro Arg Ile Arg Leu Val Thr Leu Pro Glu Val Gln Asp
65 70 75 80
Pro Pro Pro Met Glu Leu Phe Val Glu Phe Ala Glu Ser Tyr Ile Leu
85 90 95
Glu Tyr Val Lys Lys Met Val Pro Ile Ile Arg Glu Ala Leu Ser Thr
100 105 110
Leu Leu Ser Ser Arg Asp Glu Ser Gly Ser Val Arg Val Ala Gly Leu
115 120 125
Val Leu Asp Phe Phe Cys Val Pro Met Ile Asp Val Gly Asn Glu Phe
130 135 140
Asn Leu Pro Ser Tyr Ile Phe Leu Thr Cys Ser Ala Gly Phe Leu Gly
145 150 155 160
Met Met Lys Tyr Leu Pro Glu Arg His Arg Glu Ile Lys Ser Glu Phe
165 170 175
Asn Arg Ser Phe Asn Glu Glu Leu Asn Leu Ile Pro Gly Tyr Val Asn
180 185 190
Ser Val Pro Thr Lys Val Leu Pro Ser Gly Leu Phe Met Lys Glu Thr
195 200 205
Tyr Glu Pro Trp Val Glu Leu Ala Glu Arg Phe Pro Glu Ala Lys Gly
210 215 220
Ile Leu Val Asn Ser Tyr Thr Ala Leu Glu Pro Asn Gly Phe Lys Tyr
225 230 235 240
Phe Asp Arg Cys Pro Asp Asn Tyr Pro Thr Ile Tyr Pro Ile Gly Pro
245 250 255
Ile Leu Cys Ser Asn Asp Arg Pro Asn Leu Asp Leu Ser Glu Arg Asp
260 265 270
Arg Ile Leu Lys Trp Leu Asp Asp Gln Pro Glu Ser Ser Val Val Phe
275 280 285
Leu Cys Phe Gly Ser Leu Lys Ser Leu Ala Ala Ser Gln Ile Lys Glu
290 295 300
Ile Ala Gln Ala Leu Glu Leu Val Gly Ile Arg Phe Leu Trp Ser Ile
305 310 315 320
Arg Thr Asp Pro Lys Glu Tyr Ala Ser Pro Asn Glu Ile Leu Pro Asp
325 330 335
Gly Phe Met Asn Arg Val Met Gly Leu Gly Leu Val Cys Gly Trp Ala
340 345 350
Pro Gln Val Glu Ile Leu Ala His Lys Ala Ile Gly Gly Phe Val Ser
355 360 365
His Cys Gly Trp Asn Ser Ile Leu Glu Ser Leu Arg Phe Gly Val Pro
370 375 380
Ile Ala Thr Trp Pro Met Tyr Ala Glu Gln Gln Leu Asn Ala Phe Thr
385 390 395 400
Ile Val Lys Glu Leu Gly Leu Ala Leu Glu Met Arg Leu Asp Tyr Val
405 410 415
Ser Glu Tyr Gly Glu Ile Val Lys Ala Asp Glu Ile Ala Gly Ala Val
420 425 430
Arg Ser Leu Met Asp Gly Glu Asp Val Pro Arg Arg Lys Leu Lys Glu
435 440 445
Ile Ala Glu Ala Gly Lys Glu Ala Val Met Asp Gly Gly Ser Ser Phe
450 455 460
Val Ala Val Lys Arg Phe Ile Asp Gly Leu
465 470
<210> 132
<211> 1425
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 132
atgggcaaac aagaagatgc cgaactggtt attattccgt ttccgtttag cggtcatatt 60
ctggcaacca ttgaactggc aaaacgtctg attagccagg ataatccgcg tattcatacc 120
attaccattc tgtattgggg tctgccgttt attccgcagg cagataccat tgcatttctg 180
cgtagcctgg ttaaaaatga accgcgtatc cgtctggtta ccctgccgga agttcaggat 240
ccgcctccga tggaactgtt tgttgaattt gcagaaagct atatcctgga atatgtgaaa 300
aaaatggtgc cgattattcg tgaagcactg agcaccctgc tgagcagccg tgatgaaagc 360
ggtagcgttc gtgttgcagg tctggttctg gattttttct gtgttccgat gattgatgtg 420
ggcaacgaat ttaatctgcc gagctatatc tttctgacct gtagcgcagg ttttctgggt 480
atgatgaaat atctgccgga acgtcatcgt gaaatcaaaa gcgaatttaa ccgcagcttt 540
aacgaagaac tgaatctgat tccgggttat gttaatagcg ttccgaccaa agtgctgccg 600
agcggtctgt ttatgaaaga aacctatgaa ccgtgggtag aactggccga acgttttccg 660
gaagcaaaag gtattctggt taatagctat accgcactgg aaccgaatgg cttcaaatat 720
ttcgatcgtt gtccggataa ctacccgacc atttatccga ttggtccgat tctgtgtagc 780
aatgatcgtc cgaatctgga tctgagcgaa cgtgatcgta ttctgaaatg gctggatgat 840
cagccggaaa gcagcgttgt gtttctgtgc tttggtagcc tgaaaagcct ggcagcaagc 900
cagattaaag aaattgcaca ggccctggaa ctggttggta ttcgttttct gtggtcaatt 960
cgtaccgatc cgaaagaata tgcaagcccg aacgaaatcc tgccggatgg ttttatgaat 1020
cgtgttatgg gtctgggttt agtttgtggt tgggcaccgc aggttgaaat tctggcacat 1080
aaagcaattg gtggttttgt tagccattgc ggttggaata gcattctgga aagcctgcgt 1140
tttggtgtgc cgattgcaac ctggccgatg tatgcagaac agcagctgaa tgcatttacc 1200
attgtgaaag aattaggtct ggcactggaa atgcgtctgg attatgttag cgaatatggc 1260
gaaattgtca aagccgatga aattgccggt gcagttcgta gcctgatgga tggtgaagat 1320
gttccgcgtc gtaaactgaa agaaatcgca gaagcaggta aagaagcagt tatggatggc 1380
ggtagcagct ttgttgcagt taaacgtttt attgatggcc tgtaa 1425
<210> 133
<211> 456
<212> PRT
<213> P. abies
<400> 133
Met Asp Asp Gly Gly Leu Ser Trp Pro Asn Arg Ile Tyr Ala Ala Pro
1 5 10 15
Gly Val Phe Gly Cys Gly Arg Pro Gly Gln Ile Ala Tyr Met Gln Arg
20 25 30
Leu Ala Ser Ser Ala Val Gly Ala Ile Asp Phe Leu Glu Leu Pro Gly
35 40 45
Val Glu Ile Glu Gly Asp His Pro Asn Met Asn Ile Arg Thr Arg Leu
50 55 60
Ser Leu Leu Met Glu Glu Thr Lys Ile Leu Val Glu Asp Ala Leu Arg
65 70 75 80
Ser Phe Arg Phe Pro Val Cys Ala Phe Ile Ala Asp Leu Phe Ala Thr
85 90 95
Ala Met Phe Asp Val Thr Ala Lys Leu Lys Ile Pro Ser Tyr Ile Phe
100 105 110
Phe Thr Ser Ser Ala Ser Leu Leu Cys Ile Leu Leu Tyr Leu Pro Thr
115 120 125
Leu Ala Gln Glu Ile Glu Ile Ser Phe Lys Asp Val Asp Phe Pro Ile
130 135 140
Glu Val Pro Gly Leu Pro Pro Ile Pro Gly Arg Asp Leu Pro Ser His
145 150 155 160
Leu Gln Asp Arg Ser Asp Asn Val Ser Phe Asn Arg Ser Ile Gln His
165 170 175
Ser Ser Gln Leu Arg Glu Ala His Gly Ile Leu Ile Asn Thr Phe Gln
180 185 190
Asp Ile Glu Ala Glu Gln Val Lys Ala Leu Leu Glu Gly Lys Val Leu
195 200 205
Ser Ala Ala Glu Met Pro Ser Ile Tyr Pro Ile Gly Pro Ile Val Ser
210 215 220
Ser Ser Arg Leu Glu Ser Glu Ser Asp Lys Glu Glu Cys Val Glu Trp
225 230 235 240
Leu Asp Gly Gln Pro Ala Ser Ser Val Leu Phe Val Ser Phe Gly Ser
245 250 255
Arg Gly Thr Leu Ser Asp Asp Gln Ile Lys Glu Leu Ala Leu Gly Leu
260 265 270
Glu Ala Ser Gly Gln Arg Phe Leu Trp Ala Leu Leu Asn Pro Pro Pro
275 280 285
Pro Ser Ile Gln Cys Glu Asn Ser Val Ser Thr Thr Ser Ala Glu Pro
290 295 300
Asp Met Arg Leu Leu Leu Pro Glu Gly Phe Glu Asn Arg Thr Lys Asp
305 310 315 320
Arg Gly Leu Val Val His Ser Trp Val Pro Gln Ile Pro Val Leu Ser
325 330 335
His Pro Ser Thr Gly Gly Phe Leu Ser His Cys Gly Trp Asn Ser Thr
340 345 350
Leu Glu Ser Ile Leu His Gly Val Pro Leu Ile Ala Leu Pro Leu Ile
355 360 365
His Asp Gln Arg Thr Asn Ala Phe Leu Leu Val Asn Glu Ala Val Ala
370 375 380
Ile Glu Ala Lys Asn Gly Pro Asp Gly Leu Val Ser Lys Glu Glu Val
385 390 395 400
Glu Arg Val Ala Arg Glu Leu Met Glu Gly Asp Gly Gly Val Lys Ile
405 410 415
Lys Lys Arg Val Arg Lys Leu Met Glu Lys Ala Lys Asn Ala Leu Val
420 425 430
Glu Gly Gly Ser Ser Tyr Asn Ser Met Ala Thr Val Ala Ala Val Trp
435 440 445
Lys Glu Leu Asp Gly His Ser Cys
450 455
<210> 134
<211> 1371
<212> DNA
<213> P. abies
<400> 134
atggatgatg gtggtctgag ctggccgaat cgtatttatg cagcaccggg tgtttttggt 60
tgtggtcgtc cgggtcagat tgcctatatg cagcgtctgg caagcagcgc agttggtgca 120
attgattttc tggaactgcc tggtgttgaa attgaaggtg atcatccgaa tatgaatatt 180
cgtacccgtc tgagcctgct gatggaagaa accaaaattc tggttgaaga tgcactgcgt 240
agctttcgtt ttccggtttg tgcatttatt gcagacctgt ttgcaaccgc aatgtttgat 300
gttaccgcca aactgaaaat tccgagctat atctttttta ccagcagcgc aagcctgctg 360
tgtattctgc tgtatctgcc gacactggca caagaaattg aaatcagctt taaagatgtg 420
gacttcccga ttgaagttcc gggtctgcct ccgattccgg gtcgtgatct gccgagccat 480
ctgcaggatc gtagcgataa tgttagcttt aatcgtagca ttcagcatag cagccagctg 540
cgtgaagcac atggtattct gattaatacc tttcaggata tcgaagccga acaggttaaa 600
gcactgctgg aaggtaaagt tctgagcgca gcagaaatgc cgagcattta tccgattggt 660
ccgattgtta gcagcagccg tctggaaagc gaaagcgata aagaagaatg tgttgaatgg 720
ctggatggtc agcctgccag cagcgttctg tttgtgagct ttggtagccg tggcaccctg 780
agtgatgatc agattaaaga actggcactg ggtttagaag caagcggtca gcgttttctg 840
tgggcactgc tgaatccgcc tccgccaagc attcagtgtg aaaatagcgt tagcaccacc 900
agtgcagaac cggatatgcg tctgctgctg ccggaaggtt ttgaaaatcg taccaaagat 960
cgtggtctgg ttgttcatag ctgggttccg cagattccgg tgctgagcca tccgagcacc 1020
ggtggttttc tgagccattg tggttggaat agcaccctgg aaagcattct gcatggtgtt 1080
ccgctgattg cactgccgct gattcacgat cagcgtacca atgcctttct gctggttaat 1140
gaagcagttg caattgaagc aaaaaatggt ccggatggtc tggtgagcaa agaagaagtt 1200
gaacgcgttg cacgtgaatt aatggaaggt gatggtggcg tgaaaatcaa aaaacgtgtt 1260
cgtaaactga tggaaaaggc caaaaatgcc ctggtggaag gtggtagcag ctataatagc 1320
atggcaaccg ttgcagcagt ttggaaagaa ttagatggtc acagctgcta a 1371
<210> 135
<211> 484
<212> PRT
<213> 拟南芥
<400> 135
Met Asn Arg Glu Val Ser Glu Arg Ile His Ile Leu Phe Phe Pro Phe
1 5 10 15
Met Ala Gln Gly His Met Ile Pro Ile Leu Asp Met Ala Lys Leu Phe
20 25 30
Ser Arg Arg Gly Ala Lys Ser Thr Leu Leu Thr Thr Pro Ile Asn Ala
35 40 45
Lys Ile Phe Glu Lys Pro Ile Glu Ala Phe Lys Asn Gln Asn Pro Asp
50 55 60
Leu Glu Ile Gly Ile Lys Ile Phe Asn Phe Pro Cys Val Glu Leu Gly
65 70 75 80
Leu Pro Glu Gly Cys Glu Asn Ala Asp Phe Ile Asn Ser Tyr Gln Lys
85 90 95
Ser Asp Ser Gly Asp Leu Phe Leu Lys Phe Leu Phe Ser Thr Lys Tyr
100 105 110
Met Lys Gln Gln Leu Glu Ser Phe Ile Glu Thr Thr Lys Pro Ser Ala
115 120 125
Leu Val Ala Asp Met Phe Phe Pro Trp Ala Thr Glu Ser Ala Glu Lys
130 135 140
Leu Gly Val Pro Arg Leu Val Phe His Gly Thr Ser Phe Phe Ser Leu
145 150 155 160
Cys Cys Ser Tyr Asn Met Arg Ile His Lys Pro His Lys Lys Val Ala
165 170 175
Thr Ser Ser Thr Pro Phe Val Ile Pro Gly Leu Pro Gly Asp Ile Val
180 185 190
Ile Thr Glu Asp Gln Ala Asn Val Ala Lys Glu Glu Thr Pro Met Gly
195 200 205
Lys Phe Met Lys Glu Val Arg Glu Ser Glu Thr Asn Ser Phe Gly Val
210 215 220
Leu Val Asn Ser Phe Tyr Glu Leu Glu Ser Ala Tyr Ala Asp Phe Tyr
225 230 235 240
Arg Ser Phe Val Ala Lys Arg Ala Trp His Ile Gly Pro Leu Ser Leu
245 250 255
Ser Asn Arg Glu Leu Gly Glu Lys Ala Arg Arg Gly Lys Lys Ala Asn
260 265 270
Ile Asp Glu Gln Glu Cys Leu Lys Trp Leu Asp Ser Lys Thr Pro Gly
275 280 285
Ser Val Val Tyr Leu Ser Phe Gly Ser Gly Thr Asn Phe Thr Asn Asp
290 295 300
Gln Leu Leu Glu Ile Ala Phe Gly Leu Glu Gly Ser Gly Gln Ser Phe
305 310 315 320
Ile Trp Val Val Arg Lys Asn Glu Asn Gln Gly Asp Asn Glu Glu Trp
325 330 335
Leu Pro Glu Gly Phe Lys Glu Arg Thr Thr Gly Lys Gly Leu Ile Ile
340 345 350
Pro Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Lys Ala Ile Gly
355 360 365
Gly Phe Val Thr His Cys Gly Trp Asn Ser Ala Ile Glu Gly Ile Ala
370 375 380
Ala Gly Leu Pro Met Val Thr Trp Pro Met Gly Ala Glu Gln Phe Tyr
385 390 395 400
Asn Glu Lys Leu Leu Thr Lys Val Leu Arg Ile Gly Val Asn Val Gly
405 410 415
Ala Thr Glu Leu Val Lys Lys Gly Lys Leu Ile Ser Arg Ala Gln Val
420 425 430
Glu Lys Ala Val Arg Glu Val Ile Gly Gly Glu Lys Ala Glu Glu Arg
435 440 445
Arg Leu Trp Ala Lys Lys Leu Gly Glu Met Ala Lys Ala Ala Val Glu
450 455 460
Glu Gly Gly Ser Ser Tyr Asn Asp Val Asn Lys Phe Met Glu Glu Leu
465 470 475 480
Asn Gly Arg Lys
<210> 136
<211> 1455
<212> DNA
<213> 拟南芥
<400> 136
atgaatcgtg aagtgagcga acgcattcac attctgtttt ttccgtttat ggcacagggt 60
catatgattc cgattctgga tatggcaaaa ctgtttagcc gtcgtggtgc aaaaagcacc 120
ctgctgacca caccgattaa tgcaaaaatc tttgaaaaac cgatcgaggc cttcaaaaat 180
cagaatccgg atctggaaat tggcatcaag atttttaact ttccgtgcgt tgaactgggt 240
ctgccggaag gttgtgaaaa tgcagatttt atcaacagct accagaaaag cgatagcggt 300
gacctgtttc tgaaatttct gttcagcacc aaatacatga aacagcagct ggaaagcttt 360
atcgaaacca ccaaaccgag cgcactggtt gcagatatgt ttttcccgtg ggcaaccgaa 420
agcgcagaaa aactgggtgt tccgcgtctg gtttttcatg gcaccagctt ttttagcctg 480
tgttgcagct ataatatgcg cattcataaa ccgcataaaa aagttgcaac cagcagcacc 540
ccgtttgtta ttccgggtct gcctggtgat attgttatta ccgaagatca ggcaaatgtg 600
gccaaagaag aaaccccgat gggcaaattt atgaaagaag ttcgcgaaag cgaaaccaat 660
agctttggtg ttctggtgaa cagcttttat gaactggaaa gcgcatatgc cgatttttat 720
cgtagctttg ttgcaaaacg tgcctggcat attggtccgc tgagcctgag caatcgcgaa 780
ctgggtgaaa aagcgcgtcg cggtaaaaaa gcaaatatcg atgaacaaga atgcctgaaa 840
tggctggata gcaaaacacc gggtagcgtt gtttatctga gctttggtag cggcaccaat 900
tttaccaatg atcagctgct ggaaatcgca tttggtctgg aaggtagcgg tcagagcttt 960
atttgggttg ttcgcaaaaa tgaaaaccag ggcgataatg aagaatggct gcctgaaggt 1020
tttaaagaac gtaccaccgg taaaggtctg attattcctg gttgggcacc gcaggttctg 1080
atcctggatc acaaagcaat tggtggcttt gttacccatt gtggttggaa tagcgcaatt 1140
gaaggtattg cagcaggtct gccgatggtt acctggccga tgggtgcaga acagttttat 1200
aacgaaaaac tgctgacaaa agtgctgcgc attggtgtta atgttggtgc aaccgaactg 1260
gtcaaaaaag gtaaactgat tagtcgtgcc caggttgaaa aagcagttcg tgaagttatt 1320
ggtggcgaaa aagccgaaga acgtcgtctg tgggcaaaaa aacttggtga aatggcaaaa 1380
gcagcagttg aagaaggtgg tagcagttat aatgacgtga acaagtttat ggaagaactg 1440
aacggtcgca aataa 1455
<210> 137
<211> 490
<212> PRT
<213> 人工
<220>
<223> 人工
<400> 137
Met Gly Lys Gln Glu Asp Ala Glu Leu Val Ile Ile Pro Phe Pro Phe
1 5 10 15
Ser Gly His Ile Leu Ala Thr Ile Glu Leu Ala Lys Arg Leu Ile Ser
20 25 30
Gln Asp Asn Pro Arg Ile His Thr Ile Thr Ile Leu Tyr Trp Gly Leu
35 40 45
Pro Phe Ile Pro Gln Ala Asp Thr Ile Ala Phe Leu Arg Ser Leu Val
50 55 60
Lys Asn Glu Pro Arg Ile Arg Leu Val Thr Leu Pro Glu Val Gln Asp
65 70 75 80
Pro Pro Pro Met Glu Leu Phe Val Glu Phe Ala Glu Ser Tyr Ile Leu
85 90 95
Glu Tyr Val Lys Lys Met Val Pro Ile Ile Arg Glu Ala Leu Ser Thr
100 105 110
Leu Leu Ser Ser Arg Asp Glu Ser Gly Ser Val Arg Val Ala Gly Leu
115 120 125
Val Leu Asp Phe Phe Cys Val Pro Met Ile Asp Val Gly Asn Glu Phe
130 135 140
Asn Leu Pro Ser Tyr Ile Phe Leu Thr Cys Ser Ala Gly Phe Leu Gly
145 150 155 160
Met Met Lys Tyr Leu Pro Glu Arg His Arg Glu Ile Lys Ser Glu Phe
165 170 175
Asn Arg Ser Phe Asn Glu Glu Leu Asn Leu Ile Pro Gly Tyr Val Asn
180 185 190
Ser Val Pro Thr Lys Val Leu Pro Ser Gly Leu Phe Met Lys Glu Thr
195 200 205
Tyr Glu Pro Trp Val Glu Leu Ala Glu Arg Phe Pro Glu Ala Lys Gly
210 215 220
Ile Leu Val Asn Ser Tyr Thr Ala Leu Glu Pro Asn Gly Phe Lys Tyr
225 230 235 240
Phe Asp Arg Cys Pro Asp Asn Tyr Pro Thr Ile Tyr Pro Ile Gly Pro
245 250 255
Ile Leu Asn Leu Glu Asn Lys Lys Asp Asp Ala Lys Thr Asp Glu Ile
260 265 270
Met Arg Trp Leu Asn Glu Gln Pro Glu Ser Ser Val Val Phe Leu Cys
275 280 285
Phe Gly Ser Met Gly Ser Phe Asn Glu Lys Gln Val Lys Glu Ile Ala
290 295 300
Val Ala Ile Glu Arg Ser Gly His Arg Phe Leu Trp Ser Leu Arg Arg
305 310 315 320
Pro Thr Pro Lys Glu Lys Ile Glu Phe Pro Lys Glu Tyr Glu Asn Leu
325 330 335
Glu Glu Val Leu Pro Glu Gly Phe Leu Lys Arg Thr Ser Ser Ile Gly
340 345 350
Lys Val Ile Gly Trp Ala Pro Gln Met Ala Val Leu Ser His Pro Ser
355 360 365
Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Thr Leu Glu Ser
370 375 380
Met Trp Cys Gly Val Pro Met Ala Ala Trp Pro Leu Tyr Ala Glu Gln
385 390 395 400
Thr Leu Asn Ala Phe Leu Leu Val Val Glu Leu Gly Leu Ala Ala Glu
405 410 415
Ile Arg Met Asp Tyr Arg Thr Asp Thr Lys Ala Gly Tyr Asp Gly Gly
420 425 430
Met Glu Val Thr Val Glu Glu Ile Glu Asp Gly Ile Arg Lys Leu Met
435 440 445
Ser Asp Gly Glu Ile Arg Asn Lys Val Lys Asp Val Lys Glu Lys Ser
450 455 460
Arg Ala Ala Val Val Glu Gly Gly Ser Ser Tyr Ala Ser Ile Gly Lys
465 470 475 480
Phe Ile Glu His Val Ser Asn Val Thr Ile
485 490
<210> 138
<211> 1473
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 138
atgggcaaac aagaagatgc cgaactggtt attattccgt ttccgtttag cggtcatatt 60
ctggcaacca ttgaactggc aaaacgtctg attagccagg ataatccgcg tattcatacc 120
attaccattc tgtattgggg tctgccgttt attccgcagg cagataccat tgcatttctg 180
cgtagcctgg ttaaaaatga accgcgtatc cgtctggtta ccctgccgga agttcaggat 240
ccgcctccga tggaactgtt tgttgaattt gcagaaagct atatcctgga atatgtgaaa 300
aaaatggtgc cgattattcg tgaagcactg agcaccctgc tgagcagccg tgatgaaagc 360
ggtagcgttc gtgttgcagg tctggttctg gattttttct gtgttccgat gattgatgtg 420
ggcaacgaat ttaatctgcc gagctatatc tttctgacct gtagcgcagg ttttctgggt 480
atgatgaaat atctgccgga acgtcatcgt gaaatcaaaa gcgaatttaa ccgcagcttt 540
aacgaagaac tgaatctgat tccgggttat gttaatagcg ttccgaccaa agtgctgccg 600
agcggtctgt ttatgaaaga aacctatgaa ccgtgggtag aactggccga acgttttccg 660
gaagcaaaag gtattctggt taatagctat accgcactgg aaccgaatgg cttcaaatat 720
ttcgatcgtt gtccggataa ctacccgacc atttatccga ttggtccgat tctgaatctg 780
gaaaacaaaa aagatgatgc caaaaccgat gaaattatgc gctggctgaa tgaacagccg 840
gaaagcagcg ttgtgtttct gtgctttggt agcatgggta gctttaatga aaaacaggtg 900
aaagaaattg ccgtggcaat tgaacgtagt ggtcatcgtt ttctgtggtc actgcgtcgt 960
ccgacaccga aagaaaaaat tgaatttccg aaagaatatg agaacctgga agaagttctg 1020
cctgaaggct ttctgaaacg taccagcagc attggtaaag ttattggttg ggcaccgcag 1080
atggcagttc tgagccatcc gagcgttggt ggttttgtta gccattgtgg ttggaatagc 1140
accctggaaa gcatgtggtg tggtgtgccg atggcagcat ggcctctgta tgcagaacag 1200
accctgaatg cctttctgct ggttgttgaa ctgggtttag cagcagaaat tcgtatggat 1260
tatcgtaccg ataccaaagc cggttatgat ggtggtatgg aagttaccgt tgaagaaatt 1320
gaagatggca ttcgcaaact gatgagtgat ggtgaaattc gcaacaaagt gaaggatgtc 1380
aaagaaaaat cacgtgcagc agttgttgaa ggtggtagca gctatgcaag tattggcaaa 1440
ttcattgaac atgtgagcaa cgtgaccatt taa 1473
<210> 139
<211> 479
<212> PRT
<213> 番木瓜(C. papaya)
<400> 139
Met Gly Lys Pro Val Asn Asp Lys His Val Leu Val Ile Pro Phe Pro
1 5 10 15
Ala Gln Gly His Met Ile Pro Leu Leu Asp Leu Thr Gln Gln Leu Ala
20 25 30
Ile Ser Gly Leu Thr Ile Thr Ile Leu Val Thr Pro Lys Asn Leu Pro
35 40 45
Ile Leu Ser Pro Leu Leu Ala Ser His Ser Ser Ile Gln Thr Leu Leu
50 55 60
Leu Pro Phe Pro Ser His Pro Ser Ile Pro Ala Gly Ala Glu Asn Thr
65 70 75 80
Lys Asp Met Pro Ala Thr Ser Phe Phe Thr Met Met Pro Val Leu Gly
85 90 95
Gln Leu His Asp Pro Leu Val His Trp Phe Asn Thr His Pro Ser Pro
100 105 110
Pro Cys Ala Val Ile Ser Asp Ile Phe Leu Gly Trp Thr His Arg Leu
115 120 125
Ala Thr Glu Leu Gly Val Arg Arg Phe Val Phe Ser Pro Ser Gly Ala
130 135 140
Phe Ala Leu Ser Ile Ile Tyr Ser Leu Trp Arg Glu Met Pro Lys Arg
145 150 155 160
Thr Asn His Asp Asn Gln Thr Glu Val Ile Ser Phe Pro Lys Leu Pro
165 170 175
Asn Ala Pro Lys Phe Asn Trp Arg Ser Val Ser Thr Ile Tyr Gln Ser
180 185 190
Tyr Val Glu Gly Asp Pro Asp Ser Glu Phe Val Lys Gln Gly Phe Trp
195 200 205
Asp Asp Met Ala Ser Trp Gly Leu Val Ile Asn Thr Phe Thr Glu Leu
210 215 220
Glu Lys Val Tyr Leu Asp His Leu Arg Ala Glu Leu Gly His Asp Arg
225 230 235 240
Ile Trp Gly Val Gly Pro Leu His Leu Leu Ala Asp Glu Ser Ser Ser
245 250 255
Glu Pro Lys Gln Arg Gly Gly Ala Ser Ser Val Ser Val Pro Glu Leu
260 265 270
Met Thr Trp Leu Asp Ser Cys Glu Asp Arg Lys Val Val Tyr Ile Cys
275 280 285
Phe Gly Ser Gln Ala Val Leu Thr Asn Ser Gln Met Ala Ala Leu Ala
290 295 300
Ser Ala Leu Glu Lys Ser Arg Val Arg Phe Val Trp Ser Val Lys Asn
305 310 315 320
Pro Thr Arg Gly Thr Gly Asn Ser Asp Lys Asp Gly Val Ile Pro Val
325 330 335
Gly Phe Glu Asn Arg Val Glu Asp Arg Gly Arg Val Ile Lys Gly Trp
340 345 350
Ala Pro Gln Val Ser Ile Leu Asn His Arg Ala Val Gly Ala Phe Leu
355 360 365
Thr His Cys Gly Trp Asn Ser Val Phe Glu Ala Val Val Ala Gly Val
370 375 380
Pro Met Leu Ala Trp Pro Met Arg Ala Asp Gln Phe Ser Asn Ala Thr
385 390 395 400
Leu Leu Val Asp Tyr Phe Lys Val Ala Thr Lys Val Cys Glu Gly Pro
405 410 415
Gln Thr Val Pro Asp Ser Thr Glu Leu Ala Arg His Phe Val Glu Leu
420 425 430
Leu Ser Glu Asn Arg Val Glu Arg Glu Lys Ala Met Glu Leu Arg Asn
435 440 445
Ala Ala Val Lys Ala Ile Lys Asp Gly Gly Ser Ser Ala Arg Asp Leu
450 455 460
Glu Lys Leu Val Gln Gln Ile Glu Glu Leu Glu Ile Gln Ser Asn
465 470 475
<210> 140
<211> 1440
<212> DNA
<213> 番木瓜
<400> 140
atgggtaaac cggtgaatga taaacatgtt ctggttattc cgtttccggc acagggtcat 60
atgattccgc tgctggatct gacacagcag ctggcaatta gcggtctgac cattaccatt 120
ctggttaccc cgaaaaatct gccgattctg agccctctgc tggcaagcca tagcagcatt 180
cagaccctgc tgctgccgtt tccgagccat ccgagcattc cggcaggcgc agaaaatacc 240
aaagatatgc ctgcaaccag cttttttacc atgatgccgg ttctgggtca gctgcatgat 300
ccgctggttc attggtttaa tacccatccg agtccgcctt gtgcagttat tagcgatatt 360
tttcttggtt ggacccatcg tctggcaacc gaactgggtg ttcgtcgttt tgtttttagc 420
ccgagcggtg catttgcact gagcattatc tatagcctgt ggcgtgaaat gccgaaacgt 480
accaatcatg ataatcagac cgaagtgatt agctttccga aactgccgaa tgcaccgaaa 540
tttaactggc gtagcgttag caccatttat cagagctatg ttgaaggtga tccggatagc 600
gaatttgtga aacaaggttt ttgggatgat atggcaagct ggggtttagt gattaatacc 660
tttacggaac tggaaaaggt gtatctggat catctgcgtg cagaactggg tcatgatcgt 720
atttggggtg ttggtccgct gcatctgctg gccgatgaaa gcagcagcga accgaaacag 780
cgtggtggtg caagcagcgt tagcgtgccg gaactgatga cctggctgga tagctgtgaa 840
gatcgtaaag ttgtgtatat ttgctttggt agccaggcag ttctgaccaa tagccagatg 900
gcagcactgg caagcgcact ggaaaaaagc cgtgttcgct ttgtttggag cgttaaaaat 960
ccgacacgtg gcaccggtaa tagcgataaa gatggtgtta ttccggtggg ttttgaaaat 1020
cgtgtggaag atcgtggtcg tgttattaaa ggttgggcac cgcaggttag cattctgaat 1080
catcgtgcag ttggtgcatt tctgacccat tgtggttgga atagcgtttt tgaagcagtt 1140
gttgccggtg ttccgatgct ggcatggccg atgcgtgccg atcagtttag caatgcaacc 1200
ctgctggttg attatttcaa agttgcaacc aaagtttgtg aaggtccgca gaccgtgccg 1260
gatagcacag aactggcacg tcattttgtt gaactgctga gcgaaaatcg cgttgaacgt 1320
gaaaaagcaa tggaactgcg taatgcagca gtgaaagcaa ttaaagatgg cggtagcagc 1380
gcacgtgatc tggaaaaact ggttcagcag attgaagaac ttgaaatcca gagcaactaa 1440
<210> 141
<211> 479
<212> PRT
<213> 潘那利番茄
<400> 141
Met Ser Glu Asn His Pro His Val Leu Ile Phe Pro Tyr Pro Ala Gln
1 5 10 15
Gly His Met Leu Pro Leu Leu Asp Phe Thr His Gln Leu Val Asn Asn
20 25 30
Gly Val His Ile Thr Ile Leu Val Thr Pro Lys Asn Leu Pro Phe Leu
35 40 45
Asn Pro Leu Leu Ser Arg Asn Pro Ser Ile Lys Thr Leu Val Leu Pro
50 55 60
Phe Pro Ser His Pro Ser Ile Pro Ala Gly Val Glu Asn Val Lys Asp
65 70 75 80
Leu Pro Ala Asn Gly Phe Leu Ser Met Met Cys Asn Leu Gly Lys Leu
85 90 95
Arg Asp Pro Ile Leu Asp Trp Phe Gly Asn His Pro Ser Pro Pro Ser
100 105 110
Ala Ile Ile Ser Asp Met Phe Leu Gly Phe Thr His Glu Ile Ala Thr
115 120 125
Gln Leu Gly Ile Arg Arg Tyr Val Phe Ser Pro Ser Gly Ala Leu Ala
130 135 140
Leu Ser Val Val Tyr Ser Leu Trp Arg Glu Met Pro Lys Arg Lys Asp
145 150 155 160
Pro Asn Asp Glu Asn Glu Asn Phe His Phe Pro Asn Ile Pro Asn Ser
165 170 175
Pro Lys Phe Pro Phe Trp Gln Ile Ser Pro Ile Tyr Arg Ser Tyr Val
180 185 190
Glu Gly Asp Pro Ser Thr Glu Phe Ile Arg Glu Cys Tyr Leu Ala Asp
195 200 205
Ile Ala Ser His Gly Ile Val Phe Asn Thr Phe Ile Glu Leu Glu Asn
210 215 220
Val Tyr Leu Asp Tyr Leu Met Lys Tyr Leu Gly His Asn Arg Val Trp
225 230 235 240
Ser Val Gly Pro Val Leu Pro Pro Gly Glu Asp Asp Val Ser Val Gln
245 250 255
Ser Asn Arg Gly Gly Ser Ser Ser Val Leu Ala Ser Glu Ile Leu Ala
260 265 270
Trp Leu Asp Arg Cys Glu Asp His Ser Val Val Tyr Val Cys Phe Gly
275 280 285
Ser Gln Ala Val Leu Thr Asn Lys Gln Met Glu Glu Leu Ala Ile Ala
290 295 300
Leu Asp Lys Ser Gly Val His Phe Ile Leu Ser Ala Lys Arg Ala Thr
305 310 315 320
Lys Gly His Ala Ser Asn Asp Tyr Gly Val Ile Pro Ser Trp Phe Glu
325 330 335
Glu Lys Val Ala Gly Arg Gly Leu Val Val Arg Asp Trp Ala Pro Gln
340 345 350
Val Leu Ile Leu Lys His Arg Ala Ile Ala Ala Phe Leu Thr His Cys
355 360 365
Gly Trp Asn Ser Thr Leu Glu Ser Leu Ile Ala Gly Val Pro Leu Leu
370 375 380
Thr Trp Pro Met Gly Ala Asp Gln Phe Ala Asn Ala Asn Leu Leu Val
385 390 395 400
Asp Glu His Glu Val Ala Ile Arg Ala Cys Glu Gly Ala Gln Thr Val
405 410 415
Pro Asn Ser Asp Glu Leu Ala Ala Leu Leu Ala Glu Ala Val Gln Gly
420 425 430
Asn Lys Val Glu Glu Arg Arg Leu Arg Ala Ser Lys Leu Arg Lys Ile
435 440 445
Ala Ile Asn Gly Ile Lys Glu Gly Gly Asn Ser Phe Lys Glu Leu Ala
450 455 460
Ala Phe Val Lys His Leu Arg Glu Glu Ala Thr Ile Ile Glu Ala
465 470 475
<210> 142
<211> 1440
<212> DNA
<213> 潘那利番茄
<400> 142
atgagcgaaa atcatccgca tgttctgatt tttccgtatc cggcacaggg tcatatgctg 60
ccgctgctgg attttaccca tcagctggtt aataatggtg tgcatattac cattctggtg 120
accccgaaaa atctgccgtt tctgaatccg ctgctgagcc gtaatccgag cattaaaacc 180
ctggttctgc cttttccgag ccatccgagt attccggcag gcgttgaaaa tgttaaagat 240
ctgcctgcaa atggctttct gagcatgatg tgtaatctgg gtaaactgcg tgatccgatt 300
ctggattggt ttggtaatca tccgagtccg cctagcgcaa ttattagcga tatgtttctg 360
ggctttaccc atgaaattgc aacacagctg ggtattcgtc gttatgtttt tagcccgagc 420
ggtgcactgg cactgagcgt tgtttatagc ctgtggcgtg aaatgccgaa acgtaaagat 480
ccgaatgatg aaaacgagaa ctttcacttt ccgaatattc cgaacagccc gaaatttccg 540
ttttggcaga ttagcccgat ttatcgtagc tatgttgaag gtgatccgag caccgaattt 600
attcgtgaat gttatctggc agatattgcg agccatggca ttgtgtttaa cacctttatt 660
gaactggaaa acgtgtacct ggactacctg atgaaatatc tgggtcataa tcgtgtttgg 720
agcgttggtc cggttctgcc accgggtgaa gatgatgtta gcgttcagag caatcgtggt 780
ggtagcagca gcgttctggc aagcgaaatt ctggcatggc tggatcgttg tgaagatcat 840
agcgttgtgt atgtttgttt tggtagccag gcagttctga ccaataaaca aatggaagaa 900
ctggcaattg cgctggataa aagcggtgtt cattttattc tgagcgcaaa acgtgcaacc 960
aaaggtcatg caagcaatga ttatggtgtt attccgagct ggtttgaaga aaaagttgca 1020
ggtcgtggtc tggttgttcg tgattgggca cctcaggttc tgattctgaa acatcgtgca 1080
attgccgcat ttctgaccca ttgtggttgg aatagcaccc tggaaagcct gattgccggt 1140
gttcctctgc tgacctggcc gatgggtgca gatcagtttg caaatgcaaa tctgctggtt 1200
gatgaacatg aagttgcaat tcgtgcatgt gaaggtgcac agaccgttcc gaatagtgat 1260
gaactggcag cactgctggc agaagcagtt cagggtaata aagttgaaga acgtcgtctg 1320
cgtgcaagca aactgcgtaa aattgcgatt aacggtatta aagaaggtgg caacagcttt 1380
aaagagctgg cagcatttgt aaaacatctg cgtgaagaag cgaccattat tgaagcataa 1440
<210> 143
<211> 470
<212> PRT
<213> T. cacao
<400> 143
Met Asp Thr Ile Ser Ser Asn Cys Ser Ser His His Ala Val Leu Phe
1 5 10 15
Pro Phe Met Ser Lys Gly His Thr Ile Pro Ile Leu His Leu Ala Arg
20 25 30
Leu Leu Leu Arg Arg Gly Leu Ala Val Thr Val Phe Thr Thr Pro Gly
35 40 45
Asn Arg Pro Phe Ile Ala Lys Ser Leu Ala Asp Thr Ser Ala Ser Ile
50 55 60
Ile Asp Ile Asn Tyr Pro Glu Asn Ile Pro Glu Ile Pro Ala Gly Val
65 70 75 80
Glu Ser Thr Asp Ala Leu Pro Ser Ile Ser Leu Phe Val Pro Phe Cys
85 90 95
Ala Ala Thr Lys Leu Met Gln His Glu Phe Glu Arg Lys Leu Gln Ser
100 105 110
Leu Leu Pro Val Ser Phe Val Val Ser Asp Gly Phe Leu Trp Trp Thr
115 120 125
Leu Glu Ser Ala Thr Lys Phe Gly Leu Pro Arg Leu Met Phe Asn Gly
130 135 140
Met Ser Gln Tyr Ala Ser Thr Val Ser Lys Ala Val Ala Glu Asp Arg
145 150 155 160
Leu Leu Phe Gly Pro Glu Ser Asp Asp Glu Leu Ile Thr Val Thr Gln
165 170 175
Phe Pro Trp Ile Arg Val Thr Arg Asn Asp Phe Glu Pro Ile Leu Ser
180 185 190
Ser Lys Pro Asp Pro Asp Ser Pro Pro Met Arg Leu Phe Met Asp Gln
195 200 205
Val Ile Ala Ala Glu Asn Ser Lys Gly Lys Leu Val Asn Ser Phe Tyr
210 215 220
Glu Leu Glu Lys Tyr Phe Phe Asp Ser Cys Asn Leu Glu Glu Arg Leu
225 230 235 240
Lys Ala Trp Ser Val Gly Pro Leu Cys Leu Ser Glu Pro Pro Lys Val
245 250 255
Glu His Glu His Glu Pro Lys Lys Lys Pro Ser Trp Ile Lys Trp Leu
260 265 270
Asp Gln Lys Leu Asp Glu Gly Cys Ser Val Leu Tyr Val Ala Phe Gly
275 280 285
Ser Gln Ala Asp Ile Ser Ser Glu Gln Leu Lys Gln Ile Ala Thr Gly
290 295 300
Leu Glu Glu Ser Lys Val Asn Phe Leu Trp Val Val Arg Lys Lys Glu
305 310 315 320
Ser Glu Leu Gly Glu Gly Phe Glu Glu Arg Val Lys Glu Thr Gly Ile
325 330 335
Val Val Arg Glu Trp Val Asp Gln Lys Glu Ile Leu Met His Gln Ser
340 345 350
Val Gln Gly Phe Leu Ser His Cys Gly Trp Asn Ser Val Leu Glu Ser
355 360 365
Ile Cys Ala Gly Val Pro Ile Leu Ala Trp Pro Met Met Ala Asp Gln
370 375 380
Pro Leu Asn Ala Arg Met Val Val Glu Glu Ile Lys Val Gly Leu Arg
385 390 395 400
Val Glu Thr Cys Asp Gly Thr Val Lys Gly Leu Val Lys Trp Glu Gly
405 410 415
Leu Met Lys Met Val Arg Glu Leu Met Glu Gly Glu Met Gly Lys Glu
420 425 430
Val Arg Ile Lys Val Lys Glu Leu Ala Glu Leu Ala Lys Met Ala Met
435 440 445
Glu Glu Asn Thr Gly Ser Ser Trp Arg Thr Leu Asp Met Leu Ile Asn
450 455 460
Glu Phe Cys Asn Asn Lys
465 470
<210> 144
<211> 1413
<212> DNA
<213> 可加树(T. cacao)
<400> 144
atggatacca ttagcagcaa ttgtagcagc catcatgcag ttctgtttcc gtttatgagc 60
aaaggtcata ccattccgat tctgcatctg gcacgtctgc tgctgcgtcg tggtctggca 120
gttaccgttt ttaccacacc gggtaatcgt ccgtttattg caaaaagcct ggcagatacc 180
agcgcaagca ttatcgatat taactatccg gaaaacatcc cggaaattcc ggcaggcgtt 240
gaaagcaccg atgcactgcc gagcattagc ctgtttgttc cgttttgtgc agcaaccaaa 300
ctgatgcagc atgaatttga acgtaaactg cagagcctgc tgccggttag ctttgttgtt 360
agtgatggtt ttctgtggtg gaccctggaa agcgcaacaa aatttggtct gcctcgtctg 420
atgtttaatg gcatgagcca gtatgcaagc accgttagca aagcagttgc agaagatcgt 480
ctgctgtttg gtccggaaag tgatgatgaa ctgattaccg ttacacagtt tccgtggatt 540
cgtgttaccc gtaatgattt tgaaccgatt ctgagcagca aaccggatcc tgatagccct 600
ccgatgcgtc tgtttatgga tcaggttatt gcagccgaaa acagcaaagg taaactggtg 660
aatagcttct acgagctgga aaagtatttt ttcgatagct gcaatctgga agaacgtctg 720
aaagcatggt cagttggtcc gctgtgtctg agcgaaccgc ctaaagttga acatgaacac 780
gaaccgaaaa aaaagccgag ctggattaaa tggctggatc agaaactgga tgaaggttgt 840
agcgttctgt atgttgcatt tggtagccag gcagatatta gcagcgaaca gctgaaacaa 900
attgcaacag gcctggaaga aagcaaagtg aactttctgt gggttgtgcg taaaaaagaa 960
agcgaattag gtgaaggttt tgaagaacgc gttaaagaaa ccggtattgt tgttcgtgaa 1020
tgggtcgatc agaaagaaat tctgatgcac cagagcgttc agggttttct gagccattgt 1080
ggttggaata gcgtgctgga aagcatttgt gccggtgtgc cgattctggc atggccgatg 1140
atggcagatc agccgctgaa tgcacgtatg gttgttgaag aaattaaagt tggtctgcgt 1200
gtggaaacct gtgatggcac cgttaaaggt ctggttaaat gggaaggtct gatgaaaatg 1260
gttcgtgaac tgatggaagg tgaaatgggt aaagaagtgc gcatcaaagt taaagaactg 1320
gccgaactgg caaaaatggc aatggaagaa aataccggta gcagctggcg taccctggat 1380
atgctgatta atgaattctg caacaacaaa taa 1413
<210> 145
<211> 478
<212> PRT
<213> 刺天茄(S. indicum)
<400> 145
Met Asp Thr Arg Lys Arg Ser Ile Arg Ile Leu Met Phe Pro Trp Leu
1 5 10 15
Ala His Gly His Ile Ser Ala Phe Leu Glu Leu Ala Lys Ser Leu Ala
20 25 30
Lys Arg Asn Phe Val Ile Tyr Ile Cys Ser Ser Gln Val Asn Leu Asn
35 40 45
Ser Ile Ser Lys Asn Met Ser Ser Lys Asp Ser Ile Ser Val Lys Leu
50 55 60
Val Glu Leu His Ile Pro Thr Thr Ile Leu Pro Pro Pro Tyr His Thr
65 70 75 80
Thr Asn Gly Leu Pro Pro His Leu Met Ser Thr Leu Lys Arg Ala Leu
85 90 95
Asp Ser Ala Arg Pro Ala Phe Ser Thr Leu Leu Gln Thr Leu Lys Pro
100 105 110
Asp Leu Val Leu Tyr Asp Phe Leu Gln Ser Trp Ala Ser Glu Glu Ala
115 120 125
Glu Ser Gln Asn Ile Pro Ala Met Val Phe Leu Ser Thr Gly Ala Ala
130 135 140
Ala Ile Ser Phe Ile Met Tyr His Trp Phe Glu Thr Arg Pro Glu Glu
145 150 155 160
Tyr Pro Phe Pro Ala Ile Tyr Phe Arg Glu His Glu Tyr Asp Asn Phe
165 170 175
Cys Arg Phe Lys Ser Ser Asp Ser Gly Thr Ser Asp Gln Leu Arg Val
180 185 190
Ser Asp Cys Val Lys Arg Ser His Asp Leu Val Leu Ile Lys Thr Phe
195 200 205
Arg Glu Leu Glu Gly Gln Tyr Val Asp Phe Leu Ser Asp Leu Thr Arg
210 215 220
Lys Arg Phe Val Pro Val Gly Pro Leu Val Gln Glu Val Gly Cys Asp
225 230 235 240
Met Glu Asn Glu Gly Asn Asp Ile Ile Glu Trp Leu Asp Gly Lys Asp
245 250 255
Arg Arg Ser Thr Val Phe Ser Ser Phe Gly Ser Glu Tyr Phe Leu Ser
260 265 270
Ala Asn Glu Ile Glu Glu Ile Ala Tyr Gly Leu Glu Leu Ser Gly Leu
275 280 285
Asn Phe Ile Trp Val Val Arg Phe Pro His Gly Asp Glu Lys Ile Lys
290 295 300
Ile Glu Glu Lys Leu Pro Glu Gly Phe Leu Glu Arg Val Glu Gly Arg
305 310 315 320
Gly Leu Val Val Glu Gly Trp Ala Gln Gln Arg Arg Ile Leu Ser His
325 330 335
Pro Ser Val Gly Gly Phe Leu Ser His Cys Gly Trp Ser Ser Val Met
340 345 350
Glu Gly Val Tyr Ser Gly Val Pro Ile Ile Ala Val Pro Met His Leu
355 360 365
Asp Gln Pro Phe Asn Ala Arg Leu Val Glu Ala Val Gly Phe Gly Glu
370 375 380
Glu Val Val Arg Ser Arg Gln Gly Asn Leu Asp Arg Gly Glu Val Ala
385 390 395 400
Arg Val Val Lys Lys Leu Val Met Gly Lys Ser Gly Glu Gly Leu Arg
405 410 415
Arg Arg Val Glu Glu Leu Ser Glu Lys Met Arg Glu Lys Gly Glu Glu
420 425 430
Glu Ile Asp Ser Leu Val Glu Glu Leu Val Thr Val Val Arg Arg Arg
435 440 445
Glu Arg Ser Asn Leu Lys Ser Glu Asn Ser Met Lys Lys Leu Asn Val
450 455 460
Met Met Met Glu Asn Arg Glu Gly Met Leu Ser Glu Asn Ala
465 470 475
<210> 146
<211> 1437
<212> DNA
<213> 刺天茄
<400> 146
atggataccc gtaaacgtag cattcgcatt ctgatgtttc cgtggctggc acatggtcat 60
attagcgcat ttctggaact ggcaaaaagc ctggcaaaac gtaatttcgt gatttatatc 120
tgtagcagcc aggtgaatct gaacagcatt agcaaaaata tgagcagcaa agatagcatc 180
agcgtgaaac tggttgaact gcatattccg accaccattc tgcctccgcc ttatcatacc 240
accaatggtc tgccaccgca tctgatgagc accctgaaac gtgcactgga tagcgcacgt 300
ccggcattta gcaccctgct gcagacactg aaaccggatc tggttctgta tgattttctg 360
cagagctggg caagcgaaga agcagaaagc cagaatattc cggcaatggt ttttctgagt 420
accggtgcag cagcaattag ctttattatg tatcactggt ttgaaacccg tccggaagaa 480
tatccgtttc ctgcaatcta ttttcgcgaa cacgagtatg ataacttttg ccgttttaaa 540
agcagcgata gcggcaccag cgatcagctg cgtgttagcg attgtgtgaa acgtagccat 600
gatctggtgc tgattaaaac ctttcgtgaa ctggaaggtc agtatgtgga ttttctgagc 660
gatctgaccc gcaaacgttt tgttccggtt ggtccgctgg ttcaagaggt tggttgtgat 720
atggaaaatg aaggcaacga tatcatcgaa tggctggatg gtaaagatcg tcgtagcacc 780
gtttttagca gctttggtag cgaatatttt ctgtccgcca acgaaattga agaaattgca 840
tatggcctgg aactgagcgg tctgaacttt atttgggttg ttcgttttcc gcacggtgac 900
gaaaaaatca aaatcgaaga aaaactgccg gaaggtttcc tggaacgtgt tgaaggtcgt 960
ggtctggttg tggaaggttg ggcacagcag cgtcgtattc tgagccatcc gagcgttggt 1020
ggttttctgt cacattgtgg ttggagcagc gttatggaag gtgtttatag cggtgttccg 1080
attattgcag ttccgatgca tctggatcag ccgtttaatg cacgtctggt tgaagcagtt 1140
ggttttggtg aagaagttgt tcgtagccgt cagggtaatc tggatcgtgg tgaagttgca 1200
cgtgttgtta aaaaactggt tatgggtaaa agcggtgaag gtctgcgtcg tcgtgtggaa 1260
gaactgagtg aaaaaatgcg tgaaaaaggc gaagaagaaa tcgatagcct ggtagaagaa 1320
ctggttaccg ttgttcgtcg tcgcgaacgt agcaatctga aaagcgaaaa cagcatgaaa 1380
aagctgaacg tgatgatgat ggaaaaccgt gaaggtatgc tgagcgaaaa tgcataa 1437
<210> 147
<211> 477
<212> PRT
<213> 毛果杨
<400> 147
Met Glu Asp Thr Ile Val Leu Tyr Pro Ser Pro Gly Arg Gly His Leu
1 5 10 15
Phe Ser Met Val Glu Leu Gly Lys Gln Ile Leu Glu His His Pro Ser
20 25 30
Ile Ser Ile Thr Ile Ile Ile Ser Ala Met Pro Thr Glu Ser Ile Ser
35 40 45
Ile Asp Asp Pro Tyr Phe Ser Thr Leu Cys Asn Thr Asn Pro Ser Ile
50 55 60
Thr Leu Ile His Leu Pro Gln Val Ser Leu Pro Pro Asn Thr Ser Phe
65 70 75 80
Ser Pro Leu Asp Phe Val Ala Ser Phe Phe Glu Leu Pro Glu Leu Asn
85 90 95
Asn Thr Asn Leu His Gln Thr Leu Leu Asn Leu Ser Lys Ser Ser Asn
100 105 110
Ile Lys Ala Phe Ile Ile Asp Phe Phe Cys Ser Ala Ala Phe Glu Phe
115 120 125
Val Ser Ser Arg His Asn Ile Pro Ile Tyr Phe Phe Tyr Thr Thr Cys
130 135 140
Ala Ser Gly Leu Ser Met Phe Leu His Leu Pro Ile Leu Asp Lys Ile
145 150 155 160
Ile Thr Lys Ser Leu Lys Asp Leu Asp Ile Ile Ile Asp Leu Pro Gly
165 170 175
Ile Pro Lys Ile Pro Ser Lys Glu Leu Pro Pro Ala Ile Ser Asp Arg
180 185 190
Ser His Arg Val Tyr Gln Tyr Leu Val Asp Thr Ala Lys Leu Met Ile
195 200 205
Lys Ser Ala Gly Leu Ile Ile Asn Thr Phe Glu Leu Leu Glu Arg Lys
210 215 220
Ala Leu Gln Ala Ile Gln Glu Gly Lys Cys Gly Ala Pro Asp Glu Pro
225 230 235 240
Val Pro Pro Leu Phe Cys Val Gly Pro Leu Leu Thr Thr Ser Glu Ser
245 250 255
Lys Ser Glu His Glu Cys Leu Thr Trp Leu Asp Ser Gln Pro Thr Arg
260 265 270
Ser Val Leu Phe Leu Cys Phe Gly Ser Met Gly Val Phe Asn Ser Arg
275 280 285
Gln Leu Arg Glu Thr Ala Ile Gly Leu Glu Lys Ser Gly Val Arg Phe
290 295 300
Leu Trp Val Val Arg Pro Pro Leu Ala Asp Ser Gln Thr Gln Ala Gly
305 310 315 320
Arg Ser Ser Thr Pro Asn Glu Pro Cys Leu Asp Leu Leu Leu Pro Glu
325 330 335
Gly Phe Leu Glu Arg Thr Lys Asp Arg Gly Phe Leu Val Asn Ser Trp
340 345 350
Ala Pro Gln Val Glu Ile Leu Asn His Gly Ser Val Gly Gly Phe Val
355 360 365
Thr His Cys Gly Trp Asn Ser Val Leu Glu Ala Leu Cys Ala Gly Val
370 375 380
Pro Met Val Ala Trp Pro Leu Tyr Ala Glu Gln Arg Met Asn Arg Ile
385 390 395 400
Phe Leu Val Glu Glu Met Lys Val Ala Leu Ala Phe Arg Glu Ala Gly
405 410 415
Asp Asp His Phe Val Asn Ala Ala Glu Leu Glu Glu Arg Val Ile Glu
420 425 430
Leu Met Asn Ser Lys Lys Gly Glu Ala Val Arg Glu Arg Val Leu Lys
435 440 445
Leu Arg Glu Asp Ala Val Val Ala Lys Ser Asp Gly Gly Ser Ser Cys
450 455 460
Ile Ala Met Ala Lys Leu Val Asp Cys Phe Lys Lys Gly
465 470 475
<210> 148
<211> 1434
<212> DNA
<213> 毛果杨
<400> 148
atggaagata ccattgttct gtatccgagt cctggtcgtg gtcacctgtt tagcatggtt 60
gaactgggta aacaaatcct ggaacatcat ccgagcatta gcattaccat tattatcagc 120
gcaatgccga ccgaaagcat cagcattgat gatccgtatt ttagcaccct gtgtaatacc 180
aatccgagta ttaccctgat tcatctgccg caggttagcc tgcctccgaa taccagcttt 240
agtccgctgg attttgttgc cagctttttt gaactgccgg aactgaataa tacgaatctg 300
catcagaccc tgctgaatct gagcaaaagc agcaacatta aagccttcat catcgacttt 360
ttttgcagcg cagcatttga atttgttagc agccgtcata acatcccgat ctattttttc 420
tataccacct gtgcaagcgg tctgagcatg tttctgcatc tgccgattct ggataaaatc 480
attaccaaaa gcctgaagga tctggatatt atcattgatc tgcctggcat tccgaaaatt 540
ccgagcaaag aactgcctcc ggcaattagc gatcgtagcc atcgtgttta tcagtatctg 600
gttgataccg ccaaactgat gattaaaagc gcaggtctga ttatcaacac ctttgagctg 660
ctggaacgta aagcactgca ggcaattcaa gagggtaaat gtggtgcacc ggatgaaccg 720
gtgcctccgc tgttttgtgt tggtccgctg ctgaccacca gtgaaagcaa aagcgaacat 780
gaatgtctga cctggctgga tagccagccg acacgtagcg ttctgtttct gtgttttggt 840
agcatgggtg tgtttaatag ccgtcagctg cgtgaaaccg caattggtct ggaaaaaagc 900
ggtgttcgtt ttctgtgggt tgttcgtccg cctctggcag atagtcagac ccaggcaggt 960
cgtagcagca ccccgaatga accgtgtctg gatctgctgc tgccggaagg ttttctggaa 1020
cgcaccaaag atcgtggctt tctggttaat agctgggcac cgcaggttga aattctgaat 1080
catggtagcg ttggtggttt tgttacccat tgtggttgga atagcgtgct ggaagcactg 1140
tgtgccggtg ttccgatggt tgcatggcct ctgtatgcag aacagcgtat gaatcgtatt 1200
tttctggtgg aagaaatgaa agttgcactg gcatttcgtg aagccggtga tgatcatttt 1260
gttaatgcag cagaactgga agaacgtgtg attgaactga tgaatagcaa aaaaggtgaa 1320
gccgttcgtg aacgtgttct gaaactgcgt gaagatgcag ttgttgcaaa aagtgatggt 1380
ggtagcagtt gtattgcaat ggcaaaactg gttgactgct ttaaaaaggg ctaa 1434
<210> 149
<211> 467
<212> PRT
<213> 向日葵
<400> 149
Met Glu Ser Ser Thr Val Val Met Tyr Pro Ser Pro Gly Ile Gly His
1 5 10 15
Leu Val Ser Met Val Glu Leu Gly Lys Leu Ile His Thr His His Pro
20 25 30
Ser Leu Ser Val Ile Ile Leu Ile Leu Thr Ala Pro Tyr Glu Thr Gly
35 40 45
Ala Thr Gly Lys Tyr Ile Asn Thr Val Ser Ala Thr Thr Pro Ala Ile
50 55 60
Thr Phe His His Leu Pro Ala Ile Ala Leu Pro Pro Asp Phe Ser Ser
65 70 75 80
Glu Phe Ile Asp Leu Ala Phe Gly Leu Pro Glu Leu Tyr Asn Ser Val
85 90 95
Val His Asn Thr Leu Val Ala Ile Ser Gln Lys Ser Thr Ile Lys Ala
100 105 110
Val Ile Leu Asp Phe Phe Ser Asn Ala Ala Phe Gln Val Ser Thr Asn
115 120 125
Leu Ser Leu Pro Thr Tyr Tyr Phe Phe Thr Ser Gly Thr Phe Gly Leu
130 135 140
Cys Ala Phe Leu Tyr Leu Thr Thr Leu His Lys Thr Thr Ser Lys Ser
145 150 155 160
Ile Lys Asp Leu Asn Thr Leu Leu Asp Phe Pro Gly Val Pro Pro Ile
165 170 175
His Ser Ser His Met Pro Thr Ala Ile Phe Asp Arg Glu Ser Asn Ser
180 185 190
Tyr Lys Asn Phe Met Lys Thr Ser Asn Asn Met Ala Lys Cys Ser Gly
195 200 205
Ile Ile Val Asn Ser Phe Leu Glu Leu Glu Glu Arg Ala Val Ala Thr
210 215 220
Leu Arg Asp Gly Lys Cys Ile Thr Asp Gly Pro Thr Pro Pro Ile Tyr
225 230 235 240
Phe Ile Gly Pro Leu Ile Ala Ser Gly Ser Gln Val Asp Pro Asn Glu
245 250 255
Asn Glu Cys Leu Lys Trp Leu Lys Thr Gln Pro Ser Lys Ser Val Val
260 265 270
Phe Leu Cys Phe Gly Ser Met Gly Val Phe Glu Lys Glu Gln Leu Lys
275 280 285
Glu Ile Ala Val Gly Leu Glu Arg Ser Gly Gln Arg Phe Leu Trp Val
290 295 300
Val Arg Asn Pro Pro Leu Glu Ser Ser Ser Gly Ala Lys Glu Phe Glu
305 310 315 320
Leu Asp Asp Ile Leu Pro Glu Gly Phe Leu Thr Arg Thr Lys Asp Lys
325 330 335
Gly Leu Val Val Lys Asn Trp Ala Pro Gln Pro Ala Ile Leu Gly His
340 345 350
Glu Ser Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Ser Leu
355 360 365
Glu Ala Val Val Ser Gly Val Pro Met Val Ala Trp Pro Leu Tyr Ala
370 375 380
Glu Gln Gln Met Asn Arg Val Tyr Leu Val Glu Glu Ile Lys Val Ala
385 390 395 400
Leu Trp Leu Arg Met Ser Ala Asp Gly Phe Val Gly Ala Glu Ala Val
405 410 415
Glu Glu Thr Val Arg Lys Leu Met Glu Gly Glu Glu Gly Arg Ala Val
420 425 430
Arg Glu Gln Ile Leu Glu Met Ser Gly Gly Ala Lys Ala Ala Val Glu
435 440 445
Asp Gly Gly Ser Ser Arg Leu Asp Phe Leu Lys Leu Thr Arg Pro Trp
450 455 460
Thr Asp Gln
465
<210> 150
<211> 1404
<212> DNA
<213> 向日葵
<400> 150
atggaaagca gcaccgttgt tatgtatccg agtcctggta ttggtcatct ggttagcatg 60
gttgaactgg gtaaactgat tcatacccat catccgagcc tgagcgttat tattctgatt 120
ctgaccgcac cgtatgaaac cggtgcaacc ggcaaatata tcaataccgt tagcgcaacc 180
acaccggcaa ttacctttca tcatctgcct gcaattgccc tgcctccgga ttttagcagc 240
gaatttattg atctggcatt tggtctgccg gaactgtata atagcgttgt tcataatacc 300
ctggttgcca ttagccagaa aagcaccatt aaagcagtta tcctggattt ctttagcaac 360
gcagcatttc aggttagcac caatctgagc ctgccgacct attatttctt taccagcggc 420
acctttggtc tgtgtgcatt tctgtatctg accacactgc ataaaaccac gagcaaaagc 480
attaaagatc tgaataccct gctggatttt ccgggtgttc cgcctattca tagcagccat 540
atgccgaccg caatttttga tcgtgaaagc aacagctaca aaaactttat gaaaaccagc 600
aacaacatgg ccaaatgcag cggtattatt gtgaatagct ttctggaact ggaagaacgt 660
gcagttgcaa ccctgcgtga tggtaaatgt attaccgatg gtccgacacc tccgatttat 720
ttcattggtc cgctgattgc aagcggtagc caggttgatc cgaatgaaaa tgaatgtctg 780
aaatggctga aaacccagcc gagcaaatca gttgtttttc tgtgttttgg tagcatgggc 840
gtgtttgaaa aagaacagct gaaagaaatt gccgttggtc tggaacgtag cggtcagcgt 900
tttctgtggg ttgttcgtaa tccgcctctg gaaagctcaa gcggtgcaaa agaatttgaa 960
ctggatgata tcctgccgga aggttttctg acccgtacca aagataaagg tctggttgtg 1020
aaaaattggg caccgcagcc tgccattctg ggtcatgaaa gcgttggtgg ttttgttagc 1080
cattgtggtt ggaatagcag cctggaagca gttgttagcg gtgttccgat ggttgcatgg 1140
cctctgtatg cagaacagca gatgaatcgt gtttatctgg tggaagaaat taaagttgca 1200
ctgtggctgc gtatgagcgc agatggtttt gtgggtgcag aagccgttga agaaaccgtt 1260
cgcaaactga tggaaggtga agagggtcgt gcagttcgtg agcagattct ggaaatgagc 1320
ggtggtgcca aagcagcagt tgaagatggt ggtagcagcc gtctggattt cctgaaactg 1380
acccgtccgt ggaccgatca gtaa 1404
<210> 151
<211> 486
<212> PRT
<213> 猕猴桃(A. chinensis)
<400> 151
Met Ala Thr Gln Ala His Gln Pro His Phe Ile Val Phe Pro Leu Met
1 5 10 15
Ala Gln Gly His Met Ile Pro Met Ile Asp Ile Ala Lys Leu Leu Ala
20 25 30
Gln Arg Gly Val Lys Val Thr Ile Val Thr Thr Pro Leu Asn Ala Glu
35 40 45
Gln Phe Lys Thr Ile Ile Ala Arg Ala Lys Leu Ser Ile Gln Phe Leu
50 55 60
Glu Leu Gly Phe Pro Cys Lys Glu Ala Gly Leu Pro Glu Gly Cys Glu
65 70 75 80
Asn Leu Asp Lys Leu Pro Ser Phe Asp Trp Ala Ser Lys Phe Phe Val
85 90 95
Ala Thr Ser Leu Leu Lys Glu Pro Leu Glu Gln Lys Leu Gly Glu Met
100 105 110
Lys Pro Lys Pro Ser Cys Ile Ile Ser Asp Met Gly Phe Pro Trp Thr
115 120 125
Ser Asp Leu Ala Thr Lys Phe His Ile Pro Arg Leu Val Phe His Gly
130 135 140
Thr Cys Cys Phe Ser Leu Leu Cys Ser Leu Asn Val Lys Ala His Asn
145 150 155 160
Val Leu Asp Gln Val Asn Ser Asp Ser Glu Tyr Phe Val Val Pro Gly
165 170 175
Leu Pro His Lys Ile Glu Leu Thr Lys Ala Gln Leu Pro Gly Phe Asn
180 185 190
Pro Ser Ser Ser Ser Gly Leu Lys Ser Val Ser Asp Gln Ile Arg Lys
195 200 205
Ala Glu Lys Glu Val Tyr Gly Val Val Val Asn Thr Phe Glu Glu Leu
210 215 220
Glu Ala Glu Tyr Val Met Gly Tyr Lys Lys Ala Lys Gly Glu Arg Val
225 230 235 240
Trp Cys Ile Gly Pro Val Ser Met Cys Asn Lys Glu Val Leu Asp Lys
245 250 255
Ala Asp Arg Gly Lys Lys Ala Ser Ile Asp Glu His His Cys Leu Lys
260 265 270
Trp Leu Asp Ser His Asp Pro Gly Ser Val Ile Tyr Ala Cys Leu Gly
275 280 285
Ser Leu Ser Arg Leu Thr Thr Pro Gln Met Ile Glu Ile Gly Leu Gly
290 295 300
Leu Glu Glu Ser Asn Arg Pro Phe Ile Trp Val Val Arg Glu Asn Ser
305 310 315 320
Asp Gly Leu Glu Lys Trp Met Leu Glu Glu Gly Phe Glu Glu Arg Thr
325 330 335
Arg Glu Arg Gly Leu Leu Ile Arg Gly Trp Ala Pro Gln Val Leu Ile
340 345 350
Leu Ser His Pro Ser Ile Gly Ala Phe Phe Thr His Cys Gly Trp Asn
355 360 365
Ser Thr Leu Glu Gly Val Cys Ala Gly Val Pro Met Met Thr Trp Pro
370 375 380
Met Phe Ala Glu Gln Phe Cys Asn Glu Lys Leu Val Val Gln Val Leu
385 390 395 400
Arg Ile Gly Val Ser Leu Gly Val Glu Val Pro Met Arg Trp Gly Glu
405 410 415
Glu Glu Lys Val Gly Val Leu Val Lys Lys Asp Thr Val Lys Glu Ala
420 425 430
Ile Asp Glu Leu Met Asp Gly Gly Ile Glu Gly Glu Glu Arg Arg Thr
435 440 445
Arg Ala Arg Gln Leu Gly Glu Met Ala Asn Arg Ala Thr Glu Glu Ala
450 455 460
Gly Ser Ser His Leu Asn Ile Thr Met Leu Ile Gln Asp Val Met Glu
465 470 475 480
Tyr Ala Asn Ser Asp Gln
485
<210> 152
<211> 1461
<212> DNA
<213> 猕猴桃
<400> 152
atggcaaccc aggcacatca gccgcatttt attgtttttc cgctgatggc acagggtcat 60
atgattccga tgattgatat tgcaaaactg ctggcacagc gtggtgttaa agttaccatt 120
gttaccacac cgctgaatgc cgaacagttt aaaaccatta ttgcacgtgc caaactgagc 180
attcagtttc tggaactggg ttttccgtgt aaagaagcag gtctgccgga aggttgtgaa 240
aatctggata aactgccgag ctttgattgg gcaagcaaat ttttcgttgc aaccagcctg 300
ctgaaagaac cgctggaaca gaaactgggt gaaatgaaac cgaaaccgag ctgtattatt 360
agcgatatgg gctttccgtg gaccagcgat ctggcaacca aatttcatat tccgcgtctg 420
gtttttcatg gcacctgttg ttttagcctg ctgtgtagcc tgaatgttaa agcacataat 480
gttctggatc aggtgaatag cgatagcgaa tattttgttg ttccgggtct gccgcataaa 540
attgaactga ccaaagcaca gctgcctggt tttaatccga gcagcagcag cggtctgaaa 600
agcgttagcg atcagattcg taaagccgaa aaagaagttt acggcgttgt tgtgaatacc 660
tttgaagaac tggaagccga atatgtgatg ggttacaaaa aagcaaaagg tgaacgtgtt 720
tggtgtattg gtccggttag catgtgtaat aaagaggtgc tggataaagc agaccgtggt 780
aaaaaagcca gcattgatga acatcattgt ctgaaatggc tggatagcca tgatccgggt 840
agcgttattt atgcatgtct gggtagcctg agccgtctga caacaccgca gatgattgaa 900
atcggtctgg gtttagaaga aagcaaccgt ccgtttattt gggttgttcg tgaaaatagt 960
gatggcctgg aaaaatggat gctggaagaa ggttttgagg aacgtacccg tgaacgtggt 1020
ctgctgattc gtggttgggc accgcaggtt ctgattctga gccatccgag cattggtgca 1080
ttttttaccc attgtggttg gaatagcacc ctggaaggtg tttgtgccgg tgtgccgatg 1140
atgacctggc cgatgtttgc agaacagttt tgtaatgaaa aactggtggt tcaggttctg 1200
cgtattggtg ttagcctggg tgttgaagtt ccgatgcgtt ggggtgaaga agaaaaagtt 1260
ggcgttctgg ttaaaaagga tacagtgaaa gaagccattg acgaactgat ggatggtggt 1320
attgaaggtg aagaacgtcg cacccgtgca cgtcagctgg gcgaaatggc aaatcgtgca 1380
accgaagaag ccggtagcag ccatctgaat atcaccatgc tgattcagga tgttatggaa 1440
tatgccaaca gcgatcagta a 1461
<210> 153
<211> 492
<212> PRT
<213> 刺天茄
<400> 153
Met Ala Ser Gln Ser His Gln Leu His Phe Val Leu Phe Pro Leu Met
1 5 10 15
Ala Pro Gly His Met Ile Pro Met Ile Asp Ile Ala Lys Leu Leu Ala
20 25 30
Gln Arg Ser Val Leu Val Ser Val Ile Thr Thr Pro Gln Asn Ala Ser
35 40 45
Arg Phe Gly Ser Thr Val Ala Arg Ala Val Arg Ala Gly Leu Gln Ile
50 55 60
Gln Leu Val Glu Ile Arg Phe Pro Ser Val Glu Ala Gly Leu Pro Glu
65 70 75 80
Gly Cys Glu Asn Leu Asp Thr Leu Pro Ser Leu Asp Met Ala Thr Asn
85 90 95
Phe Phe Val Ala Leu Asn Leu Leu Gln Lys Glu Val Glu Gln Val Phe
100 105 110
Asp Glu Met Lys Pro Arg Pro Ser Cys Leu Ile Ser Asp Met Gly Leu
115 120 125
Pro Trp Thr Thr Gln Ile Ala Glu Lys Phe His Ile Pro Arg Ile Val
130 135 140
Phe His Gly Thr Cys Cys Phe Ser Leu Leu Cys Ser His Asn Thr Met
145 150 155 160
Ala Ser Gln Ile Leu Asp Thr Leu Asn Ser Asp Ser Asp Tyr Phe Glu
165 170 175
Val Pro Asn Leu Pro Asp Arg Ile Lys Leu Arg Lys Ser Gln Val Thr
180 185 190
Gly Ser Thr Thr Arg Lys Ser Ala Ala Trp Lys Asp Val Ala Asp Gln
195 200 205
Ile Arg Ala Ala Glu Lys Thr Ser Tyr Gly Val Val Val Asn Ser Phe
210 215 220
Gln Glu Leu Glu Ala Glu Tyr Val Lys Glu Tyr Ser Lys Val Lys Gly
225 230 235 240
Glu Lys Val Trp Cys Ile Gly Pro Val Ser Leu Cys Asn Lys Glu Ser
245 250 255
Leu Asp Leu Ala Gln Arg Gly Asn Ser Ala Ala Val Asp Glu Gln Asn
260 265 270
Cys Leu Lys Trp Leu Asp Ser Tyr Glu Pro Gly Ser Val Val Tyr Ala
275 280 285
Ser Leu Gly Ser Leu Ala Arg Leu Thr Val Gln Gln Met Thr Glu Leu
290 295 300
Ala Leu Gly Leu Glu Glu Ser Asn Arg Pro Phe Ile Trp Ala Leu Gly
305 310 315 320
Gly Asp Lys Ser Gly Ala Leu Glu Gly Trp Ile Ser Glu Asn Gly Phe
325 330 335
Glu Glu Arg Thr Lys Asn Arg Gly Leu Leu Ile Arg Gly Trp Ala Pro
340 345 350
Gln Leu Leu Ile Leu Ser His Gln Ala Thr Gly Gly Phe Leu Thr His
355 360 365
Cys Gly Trp Asn Ser Thr Val Glu Gly Ile Ser Ala Gly Val Pro Met
370 375 380
Val Thr Trp Pro Leu Phe Ala Glu Gln Phe Cys Asn Glu Lys Leu Val
385 390 395 400
Val Glu Val Leu Arg Ile Gly Val Ser Ile Gly Val Glu Val Pro Val
405 410 415
Lys Trp Gly Glu Glu Glu Lys Val Gly Val Val Val Lys Lys Asp Asp
420 425 430
Val Lys Lys Ala Leu Asp Leu Leu Met Asp Glu Glu Glu Glu Gly Lys
435 440 445
Glu Arg Arg Arg Lys Ala Arg Glu Leu Gly Lys Leu Ala Asn Lys Ala
450 455 460
Ile Glu Glu Gly Gly Ser Ser His Val Ser Met Thr Leu Leu Ile Glu
465 470 475 480
Glu Ile Met Ala Lys Ala Asn His Gly Gly Ser Thr
485 490
<210> 154
<211> 1479
<212> DNA
<213> 刺天茄
<400> 154
atggcaagcc agagccatca gctgcatttt gttctgtttc cgctgatggc accgggtcat 60
atgattccga tgattgatat tgcaaaactg ctggcacagc gtagcgttct ggttagcgtt 120
attaccacac cgcagaatgc aagccgtttt ggtagcaccg ttgcacgtgc cgttcgtgca 180
ggtctgcaga ttcagctggt tgaaattcgt tttccgagcg ttgaagccgg tctgccggaa 240
ggttgtgaaa atctggatac cctgccgagc ctggatatgg caaccaactt ttttgttgca 300
ctgaacctgc tgcagaaaga agttgaacag gttttcgatg aaatgaaacc gcgtccgagc 360
tgtctgatta gcgatatggg tctgccgtgg accacacaga ttgcagaaaa atttcatatt 420
ccgcgtatcg tgtttcatgg cacctgttgt tttagcctgc tgtgtagcca taataccatg 480
gccagccaga ttctggatac actgaatagc gatagcgatt attttgaagt tccgaatctg 540
ccggatcgta ttaaactgcg taaaagccag gttaccggta gcaccacacg taaaagcgca 600
gcatggaaag atgttgcaga tcagattcgt gcagcagaaa aaaccagcta tggtgttgtt 660
gtgaacagct ttcaagaact ggaagccgaa tatgtgaaag aatacagcaa agtgaaaggc 720
gaaaaagtgt ggtgtattgg tccggttagc ctgtgtaata aagaaagtct ggatctggcc 780
cagcgtggta atagcgcagc cgttgatgaa cagaattgtc tgaaatggct ggatagctat 840
gaaccgggta gcgttgttta tgcaagcctg ggtagcctgg cacgtctgac cgttcagcag 900
atgaccgaac tggcactggg tttagaagaa agcaatcgtc cgtttatttg ggcattaggt 960
ggtgataaaa gcggtgcact ggaaggttgg attagcgaaa atggttttga agaacgtacc 1020
aaaaatcgcg gtctgctgat tcgtggctgg gcaccgcagc tgctgatcct gagtcatcag 1080
gcaaccggtg gttttctgac ccattgtggt tggaatagca ccgtggaagg tattagtgcc 1140
ggtgttccga tggttacctg gcctctgttt gcagaacagt tttgtaatga aaaactggtg 1200
gttgaagtgc tgcgtattgg tgttagcatt ggtgtggaag ttccggttaa atggggtgaa 1260
gaagagaaag ttggcgttgt ggttaaaaaa gacgatgtga aaaaagcact ggatctgctg 1320
atggatgaag aagaagaggg taaagaacgt cgtcgtaaag cacgtgaact gggtaaactg 1380
gcaaataaag caattgaaga gggtggtagc agccatgtta gcatgaccct gctgattgaa 1440
gaaattatgg caaaagcaaa tcatggtggc agcacctaa 1479
<210> 155
<211> 458
<212> PRT
<213> 可加树
<400> 155
Met Glu Ser Lys Val Asp Gln Pro His Val Ile Val Leu Pro Tyr Pro
1 5 10 15
Ala Gln Gly His Ile Asn Pro Met Phe Gln Phe Ser Lys Arg Leu Ala
20 25 30
Ser Lys Gly Phe Lys Ala Thr Leu Ala Ile Thr Val Phe Ile Ser Asn
35 40 45
Thr Met Lys Leu Glu Ser Ser Gly Ser Val Gln Ile Asp Thr Ile Ser
50 55 60
Asp Gly Tyr Asp Ala Gly Gly Leu Ala Ser Ser Gly Gly Ile Gln His
65 70 75 80
Tyr Leu Pro Arg Leu Glu Ala Ile Gly Ser Lys Thr Leu Ala Glu Leu
85 90 95
Ile Ile Lys His Lys Arg Thr Ser Arg Pro Ile Asp Cys Ile Ile Tyr
100 105 110
Asp Ala Ala Met Pro Trp Ala Leu Asp Val Ala Lys Gln Tyr Gly Leu
115 120 125
His Gly Ala Ala Phe Phe Thr Gln Met Cys Ala Val Asn Tyr Ile Tyr
130 135 140
Tyr Asn Val His His Lys Leu Leu Asn Leu Pro Ile Cys Ser Thr Pro
145 150 155 160
Ile Ser Ile Pro Gly Leu Pro Leu Leu Gln Pro Gly Asp Leu Pro Ser
165 170 175
Phe Val Cys Ser Ser Glu Gly Ser Tyr Ile Ala Tyr Leu Gly Arg Val
180 185 190
Leu Asn Gln Phe Lys Asn Ile Asp Lys Ala Asp Phe Ile Leu Ile Asn
195 200 205
Thr Phe Tyr Lys Leu Glu Asn Glu Ala Val Glu Ser Met Ser Lys Val
210 215 220
Tyr Pro Val Leu Thr Ile Gly Pro Thr Val Pro Ser Ile Tyr Leu Asp
225 230 235 240
Lys Pro Val Glu Asn Asp Lys Ala Tyr Gly Leu Asp Leu Phe Asp Phe
245 250 255
Asn Ser Ser Thr Ser Thr Asp Trp Leu Ser Thr Lys Pro Pro Gly Ser
260 265 270
Val Ile Tyr Val Ser Phe Gly Ser Val Thr Ser Ile Ser Ser Lys Gln
275 280 285
Met Glu Glu Ile Ala Arg Gly Leu Asn Asn Ser Asn Phe Tyr Phe Leu
290 295 300
Trp Val Val Arg Ala Ser Glu Glu Ala Lys Leu Pro Lys Gly Phe Lys
305 310 315 320
Glu Glu Ser Gly Glu Lys Gly Leu Ile Val Asn Trp Ser Pro Gln Leu
325 330 335
Asp Val Leu Ser Asn Glu Ala Val Gly Cys Phe Phe Thr His Cys Gly
340 345 350
Trp Asn Ser Thr Thr Glu Ala Leu Ser Leu Gly Val Pro Met Val Ala
355 360 365
Met Pro Gln Trp Thr Asp Gln Pro Thr Val Gly Lys Tyr Ile Glu Asp
370 375 380
Val Trp Lys Val Gly Val Arg Val Lys Ile Asp Asp Val Ser Gly Ile
385 390 395 400
Val Asn Arg Glu Glu Ile Glu Ser Cys Ile Arg Gln Val Met Glu Gly
405 410 415
Glu Arg Gly Lys Glu Ile Lys Glu Asn Ala Lys Lys Trp Arg Glu Leu
420 425 430
Ala Leu Glu Ala Val Gly Glu Gly Gly Thr Ser Asp Arg Asn Ile Asp
435 440 445
Glu Phe Met Ser Lys Leu Arg Arg Thr Ala
450 455
<210> 156
<211> 1377
<212> DNA
<213> 可加树
<400> 156
atggaaagca aagttgatca gccgcatgtt attgttctgc cgtatccggc acagggtcat 60
attaatccga tgtttcagtt tagcaaacgt ctggcaagca aaggttttaa agcaaccctg 120
gcaattaccg tgtttattag caataccatg aaactggaaa gcagcggtag cgttcagatt 180
gataccatta gtgatggtta tgatgccggt ggtctggcca gcagcggtgg tattcagcat 240
tatctgcctc gtctggaagc cattggtagc aaaaccctgg ccgaactgat tatcaaacat 300
aaacgtacca gccgtccgat tgattgcatt atctatgatg cagcaatgcc gtgggcatta 360
gatgttgcaa aacagtatgg tctgcatggt gcagcatttt ttacccagat gtgtgcagtg 420
aactacatct attataacgt gcatcacaaa ctgctgaatc tgccgatttg tagcaccccg 480
attagcattc cgggtctgcc gctgctgcag cctggtgatc tgccgagctt tgtttgtagc 540
agcgaaggta gctatattgc atatctgggt cgtgttctga accagttcaa aaacattgat 600
aaagccgact tcatcctgat caacaccttc tataagctgg aaaatgaagc cgttgaaagc 660
atgagcaaag tttatccggt tctgaccatt ggtccgaccg ttccgagcat ttatctggat 720
aaaccggttg aaaacgataa agcatatggt ctggacctgt ttgattttaa cagcagcacc 780
agcaccgatt ggctgagcac caaaccgcct ggtagcgtta tttatgttag ctttggtagc 840
gtgaccagca ttagcagcaa acaaatggaa gaaattgcac gcggtctgaa taacagcaac 900
ttttatttcc tgtgggttgt tcgtgcaagc gaagaagcaa aactgccgaa aggctttaaa 960
gaagaatcag gcgaaaaagg cctgattgtt aattggagtc cgcagctgga tgttctgagc 1020
aatgaagcag ttggttgctt ttttacacat tgcggttgga atagcaccac cgaagcactg 1080
agcctgggtg ttccgatggt tgcaatgccg cagtggaccg atcagccgac cgttggcaaa 1140
tatatcgaag atgtttggaa agttggtgtg cgcgtgaaaa ttgatgatgt tagcggtatt 1200
gtgaaccgcg aagaaatcga aagctgtatt cgtcaggtta tggaaggtga acgtggcaaa 1260
gaaattaaag aaaacgccaa aaaatggcgt gaactggcac tggaagcggt tggtgaaggt 1320
ggcaccagcg atcgtaatat tgatgaattt atgagcaaac tgcgtcgcac cgcataa 1377
<210> 157
<211> 480
<212> PRT
<213> 番红花
<400> 157
Met Gly Ser Glu Gly Arg Gln Leu His Ile Phe Met Phe Pro Phe Met
1 5 10 15
Ala His Gly His Met Ile Pro Ile Val Asp Met Ala Lys Leu Phe Ala
20 25 30
Ser Arg Gly Ile Lys Ile Thr Ile Val Thr Thr Pro Leu Asn Ser Ile
35 40 45
Ser Ile Ser Lys Ser Leu His Asn Cys Ser Pro Asn Ser Leu Ile Gln
50 55 60
Leu Leu Ile Leu Lys Phe Pro Ala Ala Glu Ala Gly Leu Pro Asp Gly
65 70 75 80
Cys Glu Asn Ala Asp Ser Ile Pro Ser Met Asp Leu Leu Pro Lys Phe
85 90 95
Phe Glu Ala Val Ser Leu Leu Gln Pro Pro Phe Glu Glu Ala Leu His
100 105 110
Asn Asn Arg Pro Asp Cys Leu Ile Ser Asp Met Phe Phe Pro Trp Thr
115 120 125
Asn Asp Val Ala Asp Arg Val Gly Ile Pro Arg Leu Ile Phe His Gly
130 135 140
Thr Ser Cys Phe Ser Leu Cys Ser Ser Glu Phe Met Arg Leu His Lys
145 150 155 160
Pro Tyr Gln His Val Ser Ser Asp Thr Glu Pro Phe Thr Ile Pro Tyr
165 170 175
Leu Pro Gly Asp Ile Lys Leu Thr Lys Met Lys Leu Pro Ile Phe Val
180 185 190
Arg Glu Asn Ser Glu Asn Glu Phe Ser Lys Phe Ile Thr Lys Val Lys
195 200 205
Glu Ser Glu Ser Phe Cys Tyr Gly Val Val Val Asn Ser Phe Tyr Glu
210 215 220
Leu Glu Ala Glu Tyr Val Asp Cys Tyr Lys Asp Val Leu Gly Arg Lys
225 230 235 240
Thr Trp Thr Ile Gly Pro Leu Ser Leu Thr Asn Thr Lys Thr Gln Glu
245 250 255
Ile Thr Leu Arg Gly Arg Glu Ser Ala Ile Asp Glu His Glu Cys Leu
260 265 270
Lys Trp Leu Asp Ser Gln Lys Pro Asn Ser Val Val Tyr Val Cys Phe
275 280 285
Gly Ser Leu Ala Lys Phe Asn Ser Ala Gln Leu Lys Glu Ile Ala Ile
290 295 300
Gly Leu Glu Ala Ser Gly Lys Lys Phe Ile Trp Val Val Arg Lys Gly
305 310 315 320
Lys Gly Glu Glu Glu Glu Glu Glu Gln Asn Trp Leu Pro Glu Gly Tyr
325 330 335
Glu Glu Arg Met Glu Gly Thr Gly Leu Ile Ile Arg Gly Trp Ala Pro
340 345 350
Gln Val Leu Ile Leu Asp His Pro Ser Val Gly Gly Phe Val Thr His
355 360 365
Cys Gly Trp Asn Ser Thr Leu Glu Gly Val Ala Ala Gly Val Pro Met
370 375 380
Val Thr Trp Pro Val Gly Ala Glu Gln Phe Tyr Asn Glu Lys Leu Val
385 390 395 400
Thr Glu Val Leu Lys Thr Gly Val Gly Val Gly Val Gln Lys Trp Ala
405 410 415
Pro Gly Val Gly Asp Phe Ile Glu Ser Glu Ala Val Glu Lys Ala Ile
420 425 430
Arg Arg Ile Met Glu Lys Glu Gly Glu Glu Met Arg Asn Arg Ala Ile
435 440 445
Glu Leu Gly Lys Lys Ala Lys Trp Ala Val Gly Glu Glu Gly Ser Ser
450 455 460
Tyr Ser Asn Leu Asp Ala Leu Ile Glu Glu Leu Lys Ser Leu Ala Phe
465 470 475 480
<210> 158
<211> 1443
<212> DNA
<213> 番红花
<400> 158
atgggtagcg aaggtcgtca gctgcatatc tttatgtttc cgtttatggc acatggtcat 60
atgattccga ttgtggatat ggcaaaactg tttgcaagcc gtggtatcaa aattaccatt 120
gttaccacac cgctgaacag cattagcatt agtaaaagcc tgcataattg tagcccgaat 180
agcctgattc agctgctgat tctgaaattt ccggcagccg aagcaggtct gccggatggt 240
tgtgaaaatg cagatagcat tccgagcatg gatctgctgc cgaaattctt tgaagcagtt 300
agcctgctgc agcctccgtt tgaagaagca ctgcataaca atcgtccgga ttgtctgatt 360
agcgatatgt tttttccgtg gaccaatgat gttgcagatc gtgttggtat tccgcgtctg 420
atttttcatg gcaccagctg ttttagcctg tgtagcagcg aatttatgcg tctgcataaa 480
ccgtatcagc atgttagcag cgataccgaa ccgtttacca ttccgtatct gcctggtgat 540
attaaactga ccaaaatgaa actgccgatc tttgtgcgtg aaaacagcga aaatgaattc 600
agcaaattca tcaccaaggt gaaagaaagc gaaagctttt gctatggtgt tgtggtgaac 660
agcttttatg aactggaagc cgaatatgtg gattgctata aagatgttct gggtcgtaaa 720
acctggacca ttggtccgct gagcctgacc aataccaaaa cacaagaaat taccctgcgt 780
ggtcgtgaaa gcgcaattga tgaacatgaa tgtctgaaat ggctggatag ccagaaaccg 840
aatagcgttg tttatgtttg ctttggtagc ctggccaaat ttaacagcgc acagctgaaa 900
gaaattgcca ttggtctgga agcaagcggc aaaaaattca tttgggttgt gcgtaaaggt 960
aaaggcgaag aagaagagga agaacagaat tggctgcctg aaggttatga agaacgtatg 1020
gaaggcaccg gtctgattat tcgtggttgg gcaccgcagg ttctgattct ggatcatccg 1080
agcgttggtg gttttgttac ccattgtggt tggaatagca ccctggaagg tgttgcagcc 1140
ggtgttccga tggttacctg gcctgttggt gcagaacagt tctataatga aaaactggtt 1200
accgaggtgc tgaaaaccgg tgttggtgtg ggtgttcaga aatgggcacc tggtgttggc 1260
gattttattg aaagcgaagc agttgaaaaa gccattcgtc gcattatgga aaaagaaggt 1320
gaagaaatgc gtaaccgtgc aattgaactg ggtaaaaaag caaaatgggc agttggtgaa 1380
gaaggtagca gctatagtaa tctggatgca ctgattgaag aactgaaaag cctggccttt 1440
taa 1443
<210> 159
<211> 485
<212> PRT
<213> 毛果杨
<400> 159
Met Gly Ser Leu Gly His Gln Leu His Ile Phe Phe Leu Pro Phe Phe
1 5 10 15
Ala His Gly His Met Ile Pro Ser Val Asp Met Ala Lys Leu Phe Ala
20 25 30
Ser Arg Gly Ile Lys Thr Thr Ile Ile Thr Thr Pro Leu Asn Ala Pro
35 40 45
Phe Phe Ser Lys Thr Ile Gln Lys Thr Lys Glu Leu Gly Phe Asp Ile
50 55 60
Asn Ile Leu Thr Ile Lys Phe Pro Ala Ala Glu Ala Gly Leu Pro Glu
65 70 75 80
Gly Tyr Glu Asn Thr Asp Ala Phe Ile Phe Ser Glu Asn Ala Arg Glu
85 90 95
Met Thr Ile Lys Phe Ile Lys Ala Thr Thr Phe Leu Gln Ala Pro Phe
100 105 110
Glu Lys Val Leu Gln Glu Cys His Pro Asp Cys Ile Val Ala Asp Val
115 120 125
Phe Phe Pro Trp Ala Thr Asp Ala Ala Ala Lys Phe Gly Ile Pro Arg
130 135 140
Leu Val Phe His Gly Thr Ser Asn Phe Ala Leu Ser Ala Ser Glu Cys
145 150 155 160
Val Arg Leu Tyr Glu Pro His Lys Lys Val Ser Ser Asp Ser Glu Pro
165 170 175
Phe Val Val Pro Asp Leu Pro Gly Asp Ile Lys Leu Thr Lys Lys Gln
180 185 190
Leu Pro Asp Asp Val Arg Glu Asn Val Glu Asn Asp Phe Ser Lys Phe
195 200 205
Leu Lys Ala Ser Lys Glu Ala Glu Leu Arg Ser Phe Gly Val Val Val
210 215 220
Asn Ser Phe Tyr Glu Leu Glu Pro Ala Tyr Ala Asp Tyr Tyr Lys Lys
225 230 235 240
Val Leu Gly Arg Arg Ala Trp Asn Val Gly Pro Val Ser Leu Cys Asn
245 250 255
Arg Asp Thr Glu Asp Lys Ala Gly Arg Gly Lys Glu Thr Ser Ile Asp
260 265 270
His His Glu Cys Leu Lys Trp Leu Asp Ser Lys Lys Pro Asn Ser Val
275 280 285
Val Tyr Ile Cys Phe Gly Ser Thr Thr Asn Phe Ser Asp Ser Gln Leu
290 295 300
Lys Glu Ile Ala Ala Gly Leu Glu Ala Ser Gly Gln Gln Phe Ile Trp
305 310 315 320
Val Val Arg Arg Asn Lys Lys Gly Gln Glu Asp Lys Glu Asp Trp Leu
325 330 335
Pro Glu Gly Phe Glu Glu Arg Met Glu Gly Val Gly Leu Ile Ile Arg
340 345 350
Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Glu Ala Ile Gly Ala
355 360 365
Phe Val Thr His Cys Gly Trp Asn Ser Thr Leu Glu Gly Ile Thr Ala
370 375 380
Gly Lys Pro Met Val Thr Trp Pro Ile Phe Ala Glu Gln Phe Tyr Asn
385 390 395 400
Glu Lys Leu Val Thr Asp Val Leu Lys Thr Gly Val Gly Val Gly Val
405 410 415
Lys Glu Trp Phe Arg Val His Gly Asp His Val Lys Ser Glu Ala Val
420 425 430
Glu Lys Thr Ile Thr Gln Ile Met Val Gly Glu Glu Ala Glu Glu Met
435 440 445
Arg Ser Arg Ala Lys Lys Leu Gly Glu Thr Ala Arg Lys Ala Val Glu
450 455 460
Glu Gly Gly Ser Ser Tyr Ser Asp Phe Asn Ala Leu Ile Glu Glu Leu
465 470 475 480
Arg Trp Arg Arg Pro
485
<210> 160
<211> 1458
<212> DNA
<213> 毛果杨
<400> 160
atgggtagcc tgggtcatca gctgcatatc ttttttctgc cgttttttgc acatggccat 60
atgattccga gcgttgatat ggcaaaactg tttgcaagcc gtggtattaa aaccaccatt 120
attaccacac cgctgaacgc accgtttttt agcaaaacca ttcagaaaac caaagagctg 180
ggcttcgata ttaacatcct gaccatcaaa tttccggcag cagaagcagg tctgccggaa 240
ggttatgaaa ataccgatgc atttatcttc agcgaaaatg cacgtgagat gacgatcaaa 300
ttcattaaag caaccacctt tctgcaggca ccgtttgaaa aagttctgca agaatgtcat 360
ccggattgta ttgttgccga tgtttttttt ccgtgggcaa ccgatgcagc agcaaaattt 420
ggtattccgc gtctggtttt tcatggcacc agcaattttg cactgagcgc aagcgaatgt 480
gttcgtctgt atgaaccgca taaaaaagtt agcagcgata gcgaaccgtt tgttgttccg 540
gatctgcctg gtgatattaa actgaccaaa aaacagctgc cggatgatgt tcgtgaaaat 600
gtggaaaatg acttcagcaa attcctgaaa gcaagcaaag aagcagaact gcgtagcttt 660
ggtgttgttg tgaatagctt ttatgaactg gaaccggcat atgcggacta ctacaaaaaa 720
gtgctgggtc gtcgtgcatg gaatgttggt ccggttagcc tgtgtaatcg tgataccgaa 780
gataaagcag gtcgtggtaa agaaaccagc attgatcatc atgaatgtct gaaatggctg 840
gacagcaaaa aaccgaatag cgttgtgtat atttgctttg gtagcaccac gaattttagc 900
gatagccagc tgaaagaaat tgcagccggt ctggaagcaa gcggtcagca gtttatttgg 960
gttgttcgtc gtaacaaaaa aggccaagag gataaagaag attggctgcc tgaaggcttt 1020
gaagaacgta tggaaggtgt tggtctgatt attcgtggtt gggcaccgca ggttctgatt 1080
ctggatcatg aagcaattgg tgcatttgtt acccattgtg gttggaatag caccctggaa 1140
ggtattaccg caggtaaacc gatggttacc tggccgattt ttgcagaaca gttctataat 1200
gaaaaactgg tgaccgatgt gctgaaaacc ggtgttggtg tgggtgttaa agaatggttt 1260
cgtgttcatg gtgatcacgt taaaagcgaa gcagtggaaa aaaccattac gcagattatg 1320
gttggtgaag aggccgaaga aatgcgtagc cgtgccaaaa aactgggtga aaccgcacgt 1380
aaagcagttg aagaaggtgg tagcagctat agtgatttta atgccctgat tgaagaactg 1440
cgctggcgtc gtccgtaa 1458
<210> 161
<211> 484
<212> PRT
<213> 猕猴桃
<400> 161
Met Val Ser Lys Pro His Lys Leu His Ile Tyr Phe Phe Pro Met Ile
1 5 10 15
Ala Ser Gly His Leu Ile Pro Met Val Asp Met Ala Arg Leu Phe Ala
20 25 30
Gln Arg Gly Val Lys Ala Thr Ile Ile Leu Thr Pro Phe Asn Ala Ala
35 40 45
Leu Phe Ser Lys Thr Ile Glu Arg Asp Arg Glu Leu Gly Leu Glu Thr
50 55 60
Ser Ile Arg Leu Ile Asn Phe Pro Phe Ala Glu Val Gly Met Pro Glu
65 70 75 80
Gly Cys Glu Asn Leu Ser Ser Ile Thr Ser Pro Glu Met Phe Pro Lys
85 90 95
Ile Phe Lys Ala Thr Glu Leu Leu Gln Gln Pro Leu Glu Lys Leu Leu
100 105 110
Glu Glu Asp Arg Pro Asp Cys Leu Val Ala Asp Met Tyr Phe Pro Trp
115 120 125
Ala Thr Glu Val Ala Ser Lys His Gly Ile Pro Arg Leu Ala Phe His
130 135 140
Gly Thr Gly Ala Tyr Ala Leu Cys Val His His Val Ile Ser Gln Gln
145 150 155 160
Glu Pro Tyr Lys Asn Val Glu Ser Asp Ser Glu Val Phe Thr Val Pro
165 170 175
Asp Leu Pro Asp Thr Ile Thr Met Thr Lys Arg Gln Leu Pro Asp His
180 185 190
Ile Arg Asp Gly Thr Lys Asn His Met Glu Lys Phe Ile Glu Lys Val
195 200 205
Thr Glu Ala Glu Met Lys Ser Tyr Gly Val Leu Val Asn Ser Phe His
210 215 220
Glu Leu Glu Pro Ala Tyr Ser Glu Tyr Tyr Lys Glu Val Val Gly Arg
225 230 235 240
Arg Thr Trp His Ile Gly Pro Val Ser Leu Ser Asn Arg Asp Asn Glu
245 250 255
Asp Lys Ala Arg Arg Gly Asn Lys Thr Ser Ile Asp Glu His Glu Cys
260 265 270
Leu Ser Trp Leu Ala Ser Lys Lys Pro Asn Ser Val Leu Tyr Val Cys
275 280 285
Phe Gly Ser Leu Ser Ser Phe Ser Thr Ala Gln Leu Leu Glu Ile Ala
290 295 300
Met Gly Leu Glu Ala Ser Gly Gln Gln Phe Ile Trp Val Val Arg Lys
305 310 315 320
Asp Lys Ser Lys Glu Lys Glu Asn Glu Glu Trp Leu Pro Glu Ala Phe
325 330 335
Glu Gln Arg Leu Glu Gly Arg Gly Ile Ile Ile Arg Gly Trp Ala Pro
340 345 350
Gln Val Leu Ile Leu Asp His Glu Ser Val Gly Gly Phe Met Thr His
355 360 365
Cys Gly Trp Asn Ser Ile Leu Glu Gly Val Thr Ala Gly Val Pro Met
370 375 380
Ile Thr Trp Pro His Phe Ala Glu Gln Phe Tyr Asn Glu Lys Leu Val
385 390 395 400
Thr Asn Ile Leu Arg Val Gly Val Gly Val Gly Ala Gln Glu Trp Cys
405 410 415
Arg Trp Pro Asp Asp Cys Lys Ile Tyr Val Lys Lys Glu Asp Ile Glu
420 425 430
Lys Ala Val Ala Gln Leu Met Asp Ser Glu Glu Ala Glu Glu Thr Arg
435 440 445
Ser Arg Ala Lys Ala Leu Gly Ala Met Ala Lys Lys Ala Val Glu Lys
450 455 460
Gly Gly Ser Ser Tyr Ser Asp Leu Ser Ala Phe Leu Glu Glu Leu Glu
465 470 475 480
Leu Asn Arg Asn
<210> 162
<211> 1455
<212> DNA
<213> 猕猴桃
<400> 162
atggttagca aaccgcataa actgcacatc tattttttcc cgatgattgc aagcggtcat 60
ctgattccga tggttgatat ggcacgtctg tttgcacagc gtggtgttaa agcaaccatt 120
attctgaccc cgtttaatgc agcactgttt agcaaaacca ttgaacgtga tcgtgaactg 180
ggtttagaaa ccagcattcg tctgattaac tttccgtttg ccgaagttgg tatgccggaa 240
ggttgtgaaa atctgagcag cattaccagt ccggaaatgt ttccgaaaat ctttaaagcc 300
accgaactgc tgcaacagcc gctggaaaaa ctgctggaag aagatcgtcc ggattgtctg 360
gttgcagata tgtattttcc gtgggcaacc gaagttgcaa gcaaacatgg tattccgcgt 420
ctggcatttc atggtacagg tgcctatgca ctgtgtgttc atcatgttat tagccagcaa 480
gagccgtata aaaacgttga aagcgatagc gaagttttta ccgttccgga tctgccggat 540
accattacca tgaccaaacg tcagctgccg gatcatattc gtgatggcac caaaaatcac 600
atggaaaagt ttatcgaaaa agtgaccgaa gccgagatga aaagctatgg tgttctggtt 660
aatagctttc atgaactgga accggcatat agcgaatatt acaaagaagt tgttggtcgt 720
cgtacctggc atattggtcc ggttagcctg agcaatcgtg ataatgaaga taaagcacgt 780
cgcggtaata aaacgagcat tgatgaacat gaatgtctga gctggctggc aagcaaaaaa 840
ccgaatagcg ttctgtatgt ttgttttggt agcctgagta gctttagcac cgcacagctg 900
ttagaaattg caatgggctt agaagccagc ggtcagcagt ttatttgggt tgttcgtaaa 960
gacaaatcca aagaaaaaga aaacgaagag tggctgccgg aagcatttga acagcgtctg 1020
gaaggtcgtg gtattatcat tcgtggttgg gcaccgcagg ttctgattct ggatcatgaa 1080
agtgttggtg gttttatgac ccattgtggt tggaatagca ttctggaagg cgttaccgca 1140
ggcgttccga tgattacctg gcctcatttt gcagaacagt tctataatga aaaactggtg 1200
accaacattc tgcgtgttgg tgttggcgtt ggtgcacaag aatggtgtcg ttggcctgat 1260
gattgtaaaa tctacgtgaa aaaagaggac atcgagaaag cagttgcaca gctgatggat 1320
agtgaagaag ccgaagaaac ccgtagccgt gcaaaagcac tgggtgcaat ggcaaaaaaa 1380
gccgttgaaa aaggtggtag cagctatagc gatctgagcg cctttctgga agaactggaa 1440
ttaaatcgca actaa 1455
<210> 163
<211> 478
<212> PRT
<213> B. vulgaris
<400> 163
Met Glu Glu Gln Lys Pro His Phe Leu Leu Val Thr Phe Pro Ala Gln
1 5 10 15
Gly His Val Asn Pro Ala Leu Gln Phe Ala Lys Arg Leu Leu Arg Thr
20 25 30
Gly Ala His Val Thr Phe Ser Thr Ala Ala Ser Ala His Arg Cys Phe
35 40 45
Asp Lys Ala Lys Ile Pro Ser Gly Met Ser Phe Ala Thr Phe Ser Asp
50 55 60
Gly Tyr Asp Ala Gly Phe Arg Ala Thr Asp Gly Asp Val Leu Asp Tyr
65 70 75 80
Leu Ser Thr Phe Arg Gln Arg Gly Ala Glu Thr Leu Ala Thr Leu Leu
85 90 95
Glu Asn Ser Val Ala Glu Gly Arg Pro Val Thr Cys Leu Val Tyr Thr
100 105 110
Leu Leu Leu Pro Trp Val Ala Glu Val Ala Arg Lys Phe His Val Pro
115 120 125
Ser Ala Leu Leu Trp Ile Gln Pro Ala Thr Val Phe Asp Ile Tyr Tyr
130 135 140
Tyr Tyr Phe Asn Gly Tyr His Asp Ile Ile Tyr Asp Cys Glu Lys Asp
145 150 155 160
Pro Leu Trp Ser Leu Glu Leu Pro Asn Leu Pro Leu Lys Leu Lys Ser
165 170 175
His Asp Ile Pro Ser Phe Leu Leu Pro Ser Asn Pro Phe Leu Tyr Thr
180 185 190
Phe Ala Leu Pro Thr Phe Glu Glu Gln Met Glu Glu Leu Asp Lys Glu
195 200 205
Glu Lys Pro Lys Ile Leu Val Asn Thr Phe Glu Ala Leu Glu Val Asp
210 215 220
Ala Leu Lys Ala Ile Glu Lys Phe Lys Leu Ile Pro Ile Gly Pro Leu
225 230 235 240
Leu Pro Ser Ala Phe Leu Asn Gly Lys Asp Pro Phe Asp Lys Ser Phe
245 250 255
Gly Gly Asp Leu Phe Gln Lys Thr Lys Asn Ser Asp Tyr Met Lys Trp
260 265 270
Leu Asp Ser Gln Glu Glu Tyr Ser Ser Val Ile Tyr Val Ser Phe Gly
275 280 285
Ser Ile Ser Val Leu Ser Lys Ala Gln Met Glu Glu Leu Ala Lys Ala
290 295 300
Leu Ile Gln Ile His Arg Pro Phe Leu Trp Val Ile Arg Glu Asn Glu
305 310 315 320
Lys Asp Glu Lys Asp Leu Arg Glu Glu His Asn Glu Gly Glu Leu Ser
325 330 335
Cys Met Glu Glu Leu Lys Ala Leu Gly Leu Ile Val Pro Trp Cys Ser
340 345 350
Gln Val Glu Val Leu Ser His Pro Ser Ile Gly Cys Phe Val Thr His
355 360 365
Cys Gly Trp Asn Ser Thr Leu Glu Ser Leu Thr Cys Gly Val Pro Met
370 375 380
Val Gly Phe Pro Gln Trp Thr Asp Gln Thr Thr Asn Ser Lys Leu Ile
385 390 395 400
Glu Asp Val Trp Lys Ile Gly Val Arg Val Lys Val Ser Lys Glu Glu
405 410 415
Gly Gly Leu Val Lys Ser Glu Glu Ile Lys Arg Cys Leu Glu Val Val
420 425 430
Met Glu Ser Glu Glu Met Lys Glu Asn Ala Lys Asn Trp Lys Glu Leu
435 440 445
Ala Val Glu Ala Ala Lys Glu Gly Gly Ser Ser Asp Arg Asn Leu Lys
450 455 460
Ala Phe Met Glu Glu Leu Phe Asn Val Asp Cys Lys Lys Pro
465 470 475
<210> 164
<211> 1437
<212> DNA
<213> B. vulgaris
<400> 164
atggaagaac agaaaccgca ttttctgctg gttacctttc cggcacaggg tcatgttaat 60
ccggcactgc agtttgcaaa acgtctgctg cgtaccggtg cacatgttac ctttagcacc 120
gcagcaagcg cacatcgttg ttttgataaa gcaaaaattc cgagcggtat gagctttgca 180
acctttagtg atggttatga tgcaggtttt cgtgcaaccg atggtgatgt tctggattat 240
ctgagcacct ttcgtcagcg tggtgcagaa accctggcaa ccctgctgga aaattcagtt 300
gcagaaggtc gtccggttac ctgtctggtt tataccctgc tgctgccgtg ggttgccgaa 360
gttgcacgta aatttcatgt tccgagcgca ctgctgtgga ttcagcctgc aaccgttttt 420
gatatctatt actattattt caacggctac cacgacatca tctatgattg tgaaaaagat 480
ccgctgtggt cactggaact gccgaatctg ccgctgaaac tgaaaagcca tgatattccg 540
agctttctgc tgccgagcaa tccgtttctg tatacctttg cactgccgac ctttgaagaa 600
caaatggaag aattggacaa agaagagaag ccgaaaattc tggtgaatac atttgaagcc 660
ctggaagttg atgcactgaa agccattgaa aaattcaaac tgattccgat tggtccgctg 720
ctgcctagcg catttctgaa tggtaaagat ccgtttgata aaagctttgg tggtgacctg 780
tttcagaaaa ccaaaaacag cgattacatg aaatggctgg atagccaaga agagtatagc 840
agcgttattt atgttagctt tggtagcatt agcgttctga gcaaagcaca gatggaagag 900
ttagcaaaag cactgattca gattcatcgt ccttttctgt gggtgattcg tgaaaatgaa 960
aaagacgaga aagatctgcg cgaagaacat aatgaaggtg aactgagctg tatggaagaa 1020
ctgaaggcac tgggtctgat tgttccgtgg tgtagccagg ttgaagttct gagccatccg 1080
agcattggtt gttttgttac ccattgtggt tggaatagca ccctggaaag cctgacctgt 1140
ggtgttccga tggttggttt tccgcagtgg accgatcaga ccaccaatag taaactgatt 1200
gaagatgtgt ggaaaattgg tgtgcgtgtg aaagtgagca aagaagaagg cggtctggtt 1260
aaaagcgaag aaatcaaacg ttgtctggaa gtggttatgg aatccgaaga aatgaaagag 1320
aatgccaaga actggaaaga actggcagtt gaagcagcaa aagaaggtgg tagcagcgat 1380
cgtaatctga aagcattcat ggaagaactt ttcaacgtgg actgcaaaaa accgtaa 1437
<210> 165
<211> 450
<212> PRT
<213> P. trichocarpa
<400> 165
Met Ser Glu Ala Arg Asn Asp Leu Lys His Ile Ala Val Leu Ala Phe
1 5 10 15
Pro Val Ala Thr His Gly Pro Pro Leu Leu Ser Leu Val Arg Arg Leu
20 25 30
Ser Ala Ser Ala Ser Tyr Ala Lys Phe Ser Phe Phe Ser Thr Lys Glu
35 40 45
Ser Asn Ser Lys Leu Phe Ser Lys Glu Asp Gly Leu Glu Asn Ile Lys
50 55 60
Pro Tyr Asn Val Ser Asp Gly Leu Pro Glu Asn Tyr Asn Phe Ala Gly
65 70 75 80
Asn Leu Asp Glu Val Met Asn Tyr Phe Phe Lys Ala Thr Pro Gly Asn
85 90 95
Phe Lys Gln Ala Met Glu Val Ala Val Lys Glu Val Gly Lys Asp Phe
100 105 110
Thr Cys Ile Met Ser Asp Ala Phe Leu Trp Phe Ala Ala Asp Phe Ala
115 120 125
Gln Glu Leu His Val Pro Trp Val Pro Leu Trp Thr Ser Ser Ser Arg
130 135 140
Ser Leu Leu Leu Val Leu Glu Thr Asp Leu Val His Gln Lys Met Arg
145 150 155 160
Ser Ile Ile Asn Glu Pro Glu Asp Arg Thr Ile Asp Ile Leu Pro Gly
165 170 175
Phe Ser Glu Leu Arg Gly Ser Asp Ile Pro Lys Glu Leu Phe His Asp
180 185 190
Val Lys Glu Ser Gln Phe Ala Ala Met Leu Cys Lys Ile Gly Leu Ala
195 200 205
Leu Pro Gln Ala Ala Val Val Ala Ser Asn Ser Phe Glu Glu Leu Asp
210 215 220
Pro Asp Ala Val Ile Leu Phe Lys Ser Arg Leu Pro Lys Phe Leu Asn
225 230 235 240
Ile Gly Pro Phe Val Leu Thr Ser Pro Asp Pro Phe Met Ser Asp Pro
245 250 255
His Gly Cys Leu Glu Trp Leu Asp Lys Gln Lys Gln Glu Ser Val Val
260 265 270
Tyr Ile Ser Phe Gly Ser Val Ile Ser Leu Pro Pro Gln Glu Leu Ala
275 280 285
Glu Leu Val Glu Ala Leu Lys Glu Cys Lys Leu Pro Phe Leu Trp Ser
290 295 300
Phe Arg Gly Asn Pro Lys Glu Glu Leu Pro Glu Glu Phe Leu Glu Arg
305 310 315 320
Thr Lys Glu Lys Gly Lys Val Val Ser Trp Thr Pro Gln Leu Lys Val
325 330 335
Leu Arg His Lys Ala Ile Gly Val Phe Val Thr His Ser Gly Trp Asn
340 345 350
Ser Val Leu Asp Ser Ile Ala Gly Cys Val Pro Met Ile Cys Arg Pro
355 360 365
Phe Phe Gly Asp Gln Thr Val Asn Thr Arg Thr Ile Glu Ala Val Trp
370 375 380
Gly Thr Gly Leu Glu Ile Glu Gly Gly Arg Ile Thr Lys Gly Gly Leu
385 390 395 400
Met Lys Ala Leu Arg Leu Ile Met Ser Thr Asp Glu Gly Asn Lys Met
405 410 415
Arg Lys Lys Leu Gln His Leu Gln Gly Leu Ala Leu Asp Ala Val Gln
420 425 430
Ser Ser Gly Ser Ser Thr Lys Asn Phe Glu Thr Leu Leu Glu Val Val
435 440 445
Ala Lys
450
<210> 166
<211> 1353
<212> DNA
<213> 毛果杨
<400> 166
atgagcgaag cacgtaatga cctgaaacat attgcagttc tggcatttcc ggttgcgacc 60
catggtccgc ctctgctgag cctggttcgt cgtctgagcg caagcgcaag ctatgcaaaa 120
tttagctttt ttagcaccaa agaaagcaac agcaagctgt ttagcaaaga agatggtctg 180
gaaaacatca aaccgtataa tgttagtgat ggcctgccgg aaaattacaa ttttgcaggt 240
aatctggatg aagtgatgaa ctactttttc aaagcaaccc ctggcaactt taaacaggca 300
atggaagttg cagttaaaga ggtgggtaaa gattttacct gcattatgag tgatgccttt 360
ctgtggtttg cagcagattt tgcacaagaa ctgcatgttc cgtgggttcc gctgtggacc 420
agcagcagcc gtagcctgct gttagttctg gaaaccgatc tggttcatca gaaaatgcgt 480
agcattatta acgaaccgga agatcgcacc attgatattc tgcctggttt tagcgaactg 540
cgtggtagcg atattccgaa agaactgttt catgatgtga aagaaagcca gtttgcagcc 600
atgctgtgta aaattggtct ggcactgccg caggcagcag ttgttgcaag caatagcttt 660
gaagaactgg atccggatgc cgtgattctg tttaaaagcc gtctgccgaa atttctgaat 720
attggtccgt ttgttctgac cagtccggat ccgtttatga gcgatccgca tggttgtctg 780
gaatggctgg ataaacagaa acaagaaagc gtggtgtata ttagctttgg tagcgttatt 840
agcctgcctc cgcaagaact ggcagaactg gttgaagcac tgaaagaatg taaactgccg 900
ttcctgtggt catttcgtgg taacccgaaa gaagaactgc ctgaagaatt tctggaacgc 960
acaaaagaaa aaggtaaagt tgttagctgg acaccgcagc tgaaagttct gcgtcataaa 1020
gcaattggtg tttttgttac ccatagcggt tggaatagcg ttctggatag cattgcaggt 1080
tgtgttccga tgatttgtcg tccgtttttt ggtgatcaga ccgttaatac ccgtaccatt 1140
gaagcagttt ggggcacagg cctggaaatt gaaggtggtc gtattaccaa aggtggtctg 1200
atgaaagcac tgcgtctgat tatgagcacc gatgaaggca ataaaatgcg caaaaaactg 1260
cagcatctgc aaggtctggc cctggatgca gttcagagca gcggtagcag caccaaaaac 1320
tttgaaaccc tgctggaagt tgtggccaaa taa 1353
<210> 167
<211> 449
<212> PRT
<213> 刺天茄
<400> 167
Met Thr Leu Met Lys Lys Arg Thr Ile Ile Leu Ile Pro Tyr Pro Ala
1 5 10 15
Gln Gly His Val Thr Pro Met Leu Arg Leu Ala Ser Leu Leu Ser Asn
20 25 30
Leu Gly Leu Arg Pro Val Val Ile Thr Pro Glu Phe Ile His Arg Arg
35 40 45
Ile Ser Pro Gln Ile Asn Pro Glu Asp Gly Ile Arg Cys Leu Ser Ile
50 55 60
Thr Asp Gly Leu Asp Ala Glu Thr Pro Pro Asp Phe Phe Ser Ile Glu
65 70 75 80
Arg Ala Met Glu Glu Asn Met Pro Pro Ile Leu Glu Ala Leu Leu Arg
85 90 95
Lys Met Ile Asp Glu Glu Glu Glu Glu Gly Gly Gly Ile Ala Cys Leu
100 105 110
Val Ala Asp Leu Leu Ala Ser Trp Ala Val Asp Val Ala Arg Arg Cys
115 120 125
Gly Val Ala Ala Ala Gly Phe Trp Pro Ala Met His Ala Thr Tyr Arg
130 135 140
Leu Ile Ala Ala Ile Pro His Leu Ile Arg Thr Gly Val Ile Ser Glu
145 150 155 160
Ser Gly Cys Pro Arg Asn Pro Ser Ala Pro Ile Cys Leu Ser Ser Asn
165 170 175
Glu Pro Ile Leu Thr Pro Asn Asp Leu Pro Trp Leu Ile Gly Ser Ser
180 185 190
Ser Ala Arg Ile Ser Arg Phe Lys Phe Trp Thr Arg Thr Leu Gln Arg
195 200 205
Ala Lys Thr Leu Arg Trp Leu Leu Thr Asn Thr Phe Pro Asp Glu Cys
210 215 220
Gln Ser Arg Lys Met Thr Arg Cys Ser Asn Ala Gln Gln Val Leu Glu
225 230 235 240
Ile Gly Ser Leu Ile Met Gln Ala Leu Glu Ile Ser Thr Gly Ser Phe
245 250 255
Trp Glu Asn Asp Leu Thr Cys Leu Asp Trp Leu Asp Lys Gln Thr Met
260 265 270
Gly Ser Val Met Tyr Val Ser Phe Gly Ser Trp Val Ser Pro Ile Gly
275 280 285
Glu Ala Lys Val Lys Thr Leu Ala Leu Ser Leu Gln Ala Leu Arg Arg
290 295 300
Pro Phe Ile Trp Val Leu Gly Pro Thr Trp Arg Arg Gly Leu Pro Asp
305 310 315 320
Gly Tyr Val Lys Ser Val Ala Gly His Gly Arg Ile Val Ser Trp Ala
325 330 335
Pro Gln Leu Glu Val Leu Gln His Pro Ser Val Gly Cys Tyr Leu Thr
340 345 350
His Cys Gly Trp Asn Ser Thr Met Glu Ala Ile Gln Cys Lys Lys Pro
355 360 365
Leu Leu Cys Tyr Pro Ile Ala Gly Asp Gln Phe Leu Asn Cys Ala Tyr
370 375 380
Ile Val Asn Thr Trp Arg Ile Gly Val Lys Ile Glu Gly Phe Gly Ile
385 390 395 400
Glu Glu Val Glu Asp Gly Ile Ile Lys Val Thr Glu Asp Glu Gln Val
405 410 415
Ser Trp Arg Ile Glu Arg Leu Tyr Glu Asn Leu Tyr Gly Lys Glu Gly
420 425 430
Ser Ser Lys Ala Met Ala Asn Leu Ser Thr Phe Ile Gln Asp Leu Gly
435 440 445
Lys
<210> 168
<211> 1350
<212> DNA
<213> 刺天茄
<400> 168
atgaccctga tgaaaaaacg caccattatt ctgattccgt atccggcaca gggtcatgtt 60
accccgatgc tgcgtctggc aagcctgctg agcaatctgg gtctgcgtcc ggttgttatt 120
acaccggaat ttattcatcg tcgtattagt ccgcagatta atccggaaga tggtattcgt 180
tgtctgagca ttaccgatgg tctggatgca gaaacccctc cggatttttt cagcattgaa 240
cgtgcaatgg aagaaaacat gcctccgatt ctggaagcac tgctgcgtaa aatgattgat 300
gaagaggaag aagagggcgg aggtattgca tgtctggttg ccgatctgct ggcaagctgg 360
gcagttgatg ttgcacgtcg ttgtggtgtt gcagcagcag gtttttggcc tgcaatgcat 420
gcaacctatc gtctgattgc agcaattccg catctgattc gtaccggtgt tattagcgaa 480
agcggttgtc cgcgtaatcc gagcgcaccg atttgcctga gcagcaatga accgattctg 540
accccgaatg atctgccgtg gctgattggt agcagcagcg cacgtattag ccgtttcaaa 600
ttttggaccc gtacactgca gcgtgcaaaa accctgcgtt ggctgctgac caataccttt 660
ccggatgaat gtcagagccg caaaatgacc cgttgtagca atgcccagca ggttctggaa 720
attggtagcc tgattatgca ggcactggaa attagcaccg gtagcttttg ggaaaatgat 780
ctgacctgtc tggattggct ggataaacag accatgggta gcgttatgta tgttagcttt 840
ggtagctggg ttagcccgat tggtgaagca aaagttaaaa ccctggcact gagtctgcag 900
gccctgcgtc gtccgtttat ttgggttctg ggtccgacct ggcgtcgtgg tctgccggat 960
ggttatgtta aaagcgttgc aggtcatggt cgtattgtta gctgggcacc gcagctggaa 1020
gttctgcagc atccgagcgt tggttgttat ctgacccatt gtggttggaa tagcaccatg 1080
gaagcaattc agtgtaaaaa accactgctg tgttatccga ttgccggtga tcagtttctg 1140
aattgtgcct atattgttaa tacctggcgc attggcgtta aaattgaagg ttttggtatt 1200
gaagaggtcg aggatggtat tatcaaagtg accgaagatg aacaggttag ctggcgtatt 1260
gaacgtctgt atgaaaatct gtatggtaaa gaaggttcca gcaaagcaat ggcaaatctg 1320
agcaccttta ttcaggatct gggcaaataa 1350
<210> 169
<211> 453
<212> PRT
<213> A. duranensis
<400> 169
Met Glu Lys Glu Asn Gly Lys Ala Val His Cys Val Val Leu Ala Tyr
1 5 10 15
Pro Ala Gln Gly His Ile Asn Pro Met Ile Gln Phe Ser Lys Arg Leu
20 25 30
Leu His Glu Gly Val Lys Val Thr Leu Val Thr Thr Leu Phe Tyr Gly
35 40 45
Lys Ser Leu Glu Asn Phe Pro Pro Ser Met Ser Phe Glu Thr Ile Ser
50 55 60
Asp Gly Phe Asp Asn Gly Arg His Gly Glu Gly Leu Lys Leu Thr Val
65 70 75 80
Tyr Asn Glu Val Phe Ala Gln Arg Gly Ser Gln Thr Leu Ser Glu Val
85 90 95
Leu Glu Lys Cys Ala Ile Ser Gly Tyr Pro Val Asp Cys Ile Ile Tyr
100 105 110
Asp Ser Phe Met Pro Trp Ala Leu Asp Val Ala Lys Lys Phe Gly Ile
115 120 125
Ala Gly Ala Ser Tyr Leu Thr Gln Asn Met Pro Val Asn Ser Val Tyr
130 135 140
Tyr His Val His Ile Gly Lys Leu Arg Ala Pro Leu Thr Glu Asp Glu
145 150 155 160
Ile Leu Ile Pro Met Leu Pro Lys Leu Gln His Arg Asp Met Pro Ser
165 170 175
Phe Phe Leu Ser Tyr Gln Glu Asp Pro Ala Phe Leu Glu Met Leu Val
180 185 190
Glu Gln Phe Ser Asn Ile His Glu Ala Asp Trp Val Leu Cys Asn Ala
195 200 205
Phe Tyr Glu Leu Glu Lys Glu Val Ile Asp Trp Thr Thr Lys Ile Trp
210 215 220
Pro Lys Phe Arg Thr Ile Gly Pro Ser Ile Pro Ser Met Phe Leu Asp
225 230 235 240
Lys Arg Leu Lys Asp Asp Glu Glu Tyr Gly Val Thr Gln Phe Lys Ser
245 250 255
Glu Glu Cys Met Asp Trp Leu Asp Lys Lys Ala Lys Gly Ser Val Leu
260 265 270
Tyr Val Ser Phe Gly Ser Leu Val Pro Leu Asp Glu Glu Gln Ile Arg
275 280 285
Glu Val Ala Tyr Gly Leu Arg Asp Ser Gly Arg Tyr Phe Leu Trp Val
290 295 300
Val Arg Ala Ser Glu Glu Ala Lys Leu Pro Lys Asp Phe Ala Lys Asn
305 310 315 320
Ser Glu Lys Gly Leu Val Val Thr Trp Cys Ser Gln Leu Lys Val Leu
325 330 335
Ser His Glu Ala Val Gly Cys Phe Val Thr His Cys Gly Trp Asn Ser
340 345 350
Thr Leu Glu Ala Leu Ser Leu Gly Val Pro Val Ile Ala Val Pro Gln
355 360 365
Trp Ser Asp Gln Ala Thr Asn Ala Lys Tyr Leu Val Asp Val Trp Lys
370 375 380
Val Gly Ile Arg Pro Val Val Asp Glu Lys Lys Ile Met Arg Lys Glu
385 390 395 400
Ala Leu Glu Asp Cys Ile Lys Glu Leu Met Glu Ser Asp Lys Gly Lys
405 410 415
Glu Ile Arg Ile Asn Ala Val Lys Leu Lys Asn Leu Ala Ile Glu Ala
420 425 430
Val Ser Glu Gly Gly Ser Ser Asn Lys Asn Ile Ile Glu Phe Val Asn
435 440 445
Ser Leu Lys Gly Tyr
450
<210> 170
<211> 1362
<212> DNA
<213> A. duranensis
<400> 170
atggaaaaag aaaatggcaa agccgttcat tgtgttgttc tggcatatcc ggcacagggt 60
catattaatc cgatgattca gtttagcaaa cgcctgctgc atgaaggtgt taaagttacc 120
ctggttacca cactgtttta tggtaaaagc ctggaaaact ttccgcctag catgagcttt 180
gaaaccatta gtgatggttt tgataatggc cgtcatggtg aaggtctgaa actgaccgtt 240
tataatgaag tttttgcaca gcgtggtagt cagaccctga gcgaagttct ggaaaaatgt 300
gcaattagcg gttatccggt tgattgcatt atctatgata gctttatgcc gtgggcatta 360
gatgtggcca aaaaattcgg tattgccggt gcaagctatc tgacccagaa tatgccggtt 420
aatagcgtgt attatcatgt gcatattggc aaactgcgtg caccgctgac cgaagatgaa 480
attctgattc cgatgctgcc gaaactgcag catcgtgata tgccgagctt ttttctgagc 540
tatcaagaag atcctgcctt tctggaaatg ctggttgaac agttttccaa cattcatgaa 600
gcagattggg ttctgtgcaa cgcattctat gaacttgaaa aagaagtgat cgactggacc 660
accaaaatct ggcctaaatt tcgtaccatt ggtccgagca ttccgagtat gtttctggat 720
aaacgtctga aagatgatga agaatatggc gtgacccagt ttaaaagcga agaatgtatg 780
gattggctgg acaaaaaagc aaaaggtagc gttctgtatg ttagctttgg tagcctggtt 840
ccgctggatg aagaacaaat tcgtgaagtt gcatatggtc tgcgtgatag cggtcgttat 900
tttctgtggg ttgttcgtgc cagcgaagaa gcaaaactgc cgaaagattt tgccaaaaac 960
agcgaaaaag gtctggttgt tacctggtgt agccagctga aagttctgag ccatgaagcc 1020
gttggttgtt ttgttaccca ttgtggttgg aatagcaccc tggaagcact gagcctgggt 1080
gttccggtta ttgccgttcc gcagtggtca gatcaggcaa ccaatgcaaa atatctggtt 1140
gatgtttgga aagtgggtat tcgtccggtt gttgatgaga aaaaaatcat gcgtaaagag 1200
gccctggaag attgtattaa agaactgatg gaaagcgaca aaggcaaaga aattcgtatt 1260
aatgccgtga agctgaaaaa cctggcaatt gaagcagtta gcgaaggtgg tagcagcaac 1320
aaaaacatta tcgaatttgt gaacagcctg aaaggctatt aa 1362
<210> 171
<211> 468
<212> PRT
<213> 木瓜(C. sinensis)
<400> 171
Met Glu Asn Ile Glu Lys Lys Ala Ala Ser Cys Arg Leu Val His Cys
1 5 10 15
Leu Val Leu Ser Tyr Pro Ala Gln Gly His Ile Asn Pro Leu Leu Gln
20 25 30
Phe Ala Lys Arg Leu Asp His Lys Gly Leu Lys Val Thr Leu Val Thr
35 40 45
Thr Cys Phe Ile Ser Lys Ser Leu His Arg Asp Ser Ser Ser Ser Ser
50 55 60
Thr Ser Ile Ala Leu Glu Ala Ile Ser Asp Gly Tyr Asp Glu Gly Gly
65 70 75 80
Ser Ala Gln Ala Glu Ser Ile Glu Ala Tyr Leu Glu Lys Phe Trp Gln
85 90 95
Ile Gly Pro Arg Ser Leu Cys Glu Leu Val Glu Glu Met Asn Gly Ser
100 105 110
Gly Val Pro Val Asp Cys Ile Val Tyr Asp Ser Phe Leu Pro Trp Ala
115 120 125
Leu Asp Val Ala Lys Lys Phe Gly Leu Val Gly Ala Ala Phe Leu Thr
130 135 140
Gln Ser Cys Ala Val Asp Cys Ile Tyr Tyr His Val Asn Lys Gly Leu
145 150 155 160
Leu Met Leu Pro Leu Pro Asp Ser Gln Leu Leu Leu Pro Gly Met Pro
165 170 175
Pro Leu Glu Pro His Asp Met Pro Ser Phe Val Tyr Asp Leu Gly Ser
180 185 190
Tyr Pro Ala Val Ser Asp Met Val Val Lys Tyr Gln Phe Asp Asn Ile
195 200 205
Asp Lys Ala Asp Trp Val Leu Cys Asn Thr Phe Tyr Glu Leu Glu Glu
210 215 220
Glu Val Ala Glu Trp Leu Gly Lys Leu Trp Ser Leu Lys Thr Ile Gly
225 230 235 240
Pro Thr Val Pro Ser Leu Tyr Leu Asp Lys Gln Leu Glu Asp Asp Lys
245 250 255
Asp Tyr Gly Phe Ser Met Phe Lys Pro Asn Asn Glu Ser Cys Ile Lys
260 265 270
Trp Leu Asn Asp Arg Ala Lys Gly Ser Val Val Tyr Val Ser Phe Gly
275 280 285
Ser Tyr Ala Gln Leu Lys Val Glu Glu Met Glu Glu Leu Ala Trp Gly
290 295 300
Leu Lys Ala Thr Asn Gln Tyr Phe Leu Trp Val Val Arg Glu Ser Glu
305 310 315 320
Gln Ala Lys Leu Pro Glu Asn Phe Ser Asp Glu Thr Ser Gln Lys Gly
325 330 335
Leu Val Val Asn Trp Cys Pro Gln Leu Glu Val Leu Ala His Glu Ala
340 345 350
Thr Gly Cys Phe Leu Thr His Cys Gly Trp Asn Ser Thr Met Glu Ala
355 360 365
Leu Ser Leu Gly Val Pro Met Val Ala Met Pro Gln Trp Ser Asp Gln
370 375 380
Ser Thr Asn Ala Lys Tyr Ile Met Asp Val Trp Lys Thr Gly Leu Lys
385 390 395 400
Val Pro Ala Asp Glu Lys Gly Ile Val Arg Arg Glu Ala Ile Ala His
405 410 415
Cys Ile Arg Glu Ile Leu Glu Gly Glu Arg Gly Lys Glu Ile Arg Gln
420 425 430
Asn Ala Gly Glu Trp Ser Asn Phe Ala Lys Glu Ala Val Ala Lys Gly
435 440 445
Gly Ser Ser Asp Lys Asn Ile Asp Asp Phe Val Ala Asn Leu Ile Ser
450 455 460
Ser Lys Ser Phe
465
<210> 172
<211> 1407
<212> DNA
<213> 木瓜
<400> 172
atggaaaaca tcgagaaaaa agcagcaagc tgtcgtctgg ttcattgtct ggttctgagc 60
tatccggcac agggtcatat taatccgctg ctgcagtttg caaaacgtct ggatcataaa 120
ggtctgaaag ttaccctggt taccacctgt tttattagca aaagcctgca tcgtgatagc 180
agcagcagct caaccagcat tgcactggaa gcaattagtg atggttatga tgaaggtggt 240
agcgcacagg cagaaagcat tgaagcatat ctggaaaaat tctggcagat tggtccgcgt 300
agcctgtgtg aactggttga agaaatgaat ggtagcggtg ttccggttga ttgcattgtt 360
tatgatagtt ttctgccgtg ggcattagat gtggccaaaa aattcggtct ggttggtgca 420
gcatttctga cccagagctg tgcagttgat tgtatctatt atcatgtgaa caaaggcctg 480
ctgatgctgc cgctgccgga ttcacagctg ctgttaccgg gtatgcctcc gctggaaccg 540
catgatatgc cgagctttgt gtatgatctg ggtagttatc cggcagttag cgatatggtt 600
gtgaaatatc agttcgacaa catcgataaa gcagattggg ttctgtgcaa caccttttat 660
gaactggaag aagaggttgc agaatggctg ggtaaactgt ggtcactgaa aaccattggt 720
ccgaccgttc cgagcctgta tctggataaa cagctggaag atgataaaga ttatggcttt 780
agcatgttta aaccgaacaa cgagagctgc attaaatggc tgaatgatcg tgcaaaaggt 840
agcgttgttt atgttagctt tggtagctat gcacagctga aagtggaaga aatggaagaa 900
ctggcatggg gactgaaagc aaccaatcag tattttctgt gggttgttcg tgaaagcgaa 960
caggcaaaac tgcctgaaaa ctttagtgat gaaaccagcc agaaaggtct ggtggttaat 1020
tggtgtccgc aactggaagt tctggcacat gaagccaccg gttgttttct gacacattgt 1080
ggttggaata gcaccatgga agcactgagc ctgggtgttc cgatggttgc aatgccgcag 1140
tggtcagatc agagcaccaa tgccaaatat atcatggatg tttggaaaac aggcctgaaa 1200
gttccggcag atgaaaaagg tattgttcgt cgtgaagcaa ttgcccattg tattcgtgaa 1260
attctggaag gtgaacgcgg taaagaaatt cgtcagaatg ccggtgaatg gtccaatttt 1320
gccaaagaag cagttgcaaa aggcggtagc agcgataaaa acattgatga ttttgtggcc 1380
aacctgatca gcagcaaatc cttttaa 1407
<210> 173
<211> 473
<212> PRT
<213> A. duranensis
<400> 173
Met Glu Ser Lys Thr Ile Arg Ile Ala Leu Val Ser Ala Pro Val Tyr
1 5 10 15
Ser His Leu Arg Ser Ile Leu Glu Phe Ala Lys Arg Leu Ile Arg Phe
20 25 30
Tyr Gln Asp Leu His Val Thr Cys Leu Val Pro Ile Asn Gly Ser Pro
35 40 45
Cys Asn Lys Thr Lys Ala Leu Leu Gln Ser Leu Pro Pro Thr Ile Asp
50 55 60
Tyr Ile Phe Val Ser Pro Lys Asn Leu Glu Asp Glu Val Gln Asp Thr
65 70 75 80
His Pro Ala Phe Leu Val Arg Thr Leu Ile Thr Arg Ser Leu Pro Leu
85 90 95
Ile His Asp Glu Val Lys Lys Leu Ile Ser Lys Ser Arg Leu Ile Ala
100 105 110
Ile Ile Ser Asp Gly Ile Ile Thr Gln Val Leu Glu Leu Val Lys Asp
115 120 125
Leu Asn Val Leu Ser Tyr Thr Tyr Phe Pro Ser Ser Ala Met Leu Leu
130 135 140
Ala Leu Cys Leu Tyr Ser Glu Asn Leu Asp Glu Thr Thr Thr Ser Glu
145 150 155 160
Tyr Lys Asp Leu Leu Glu Pro Ile Lys Ile Pro Gly Cys Ile Pro Val
165 170 175
Gln Gly Ser Asp Leu Pro Asp Pro Phe Asn Asp Arg Thr Ser Glu Thr
180 185 190
Tyr Lys Glu Phe Leu Glu Gly Ser Arg Arg Phe Phe Leu Ala Asp Gly
195 200 205
Ile Leu Val Asn Thr Phe Phe Asp Leu Glu Ala Ser Thr Ile Lys Glu
210 215 220
Leu Gln Glu Gln Glu Arg Arg Gly Ile Val Pro Ser Ile His Ala Ile
225 230 235 240
Gly Pro Phe Val Gln His Glu Ser Ser Met Ile Glu Gly Asn Asp Asn
245 250 255
Asn Thr Leu Glu Cys Leu Asn Trp Leu Asp Lys Gln Gln Glu Asn Ser
260 265 270
Val Leu Tyr Val Ser Phe Gly Ser Gly Gly Thr Ile Ser His Lys Gln
275 280 285
Ile Ile Glu Leu Ala Leu Gly Leu Glu Leu Ser Gly Gln Lys Phe Leu
290 295 300
Trp Leu Leu Lys Pro Pro Ser Lys Phe Asp Ile Ile Phe Asp Phe Gly
305 310 315 320
His Phe Ser Glu Asp Pro Leu Lys Tyr Leu Pro Ser Gly Phe Leu Glu
325 330 335
Arg Thr Lys Glu Gln Gly Ile Ile Val Pro Tyr Trp Ala Pro Gln Ile
340 345 350
Lys Ile Leu Gly His Ala Ala Ile Gly Gly Tyr Leu Cys His Cys Gly
355 360 365
Trp Asn Ser Ile Leu Glu Ser Val Ala His Gly Ile Pro Met Ile Ala
370 375 380
Trp Pro Leu Phe Ala Glu Gln Arg Met Asn Ala Ala Leu Phe Cys Asn
385 390 395 400
Gly Leu Lys Val Ala Ile Arg Ala Lys Val Asn Glu Met Gly Ile Val
405 410 415
Glu Arg Gly Glu Val Ala Lys Val Ile Lys Asn Leu Met Ile Gly Asp
420 425 430
Glu Gly Lys Glu Ile Arg Gln Arg Met Arg Glu Leu Lys Gly Ser Ala
435 440 445
Glu Asp Ala Ile Asn Glu Gly Gly Ser Ser Thr Arg Thr Leu Thr Gln
450 455 460
Leu Val Gln Lys Trp Lys Asn Leu Glu
465 470
<210> 174
<211> 1422
<212> DNA
<213> A. duranensis
<400> 174
atggaaagca aaaccattcg tattgcactg gttagcgcac cggtttatag ccatctgcgt 60
agcattctgg aatttgcaaa acgtctgatt cgcttctatc aggatctgca tgttacctgt 120
ctggttccga ttaatggtag cccgtgtaat aaaaccaaag cactgctgca gagcctgcct 180
ccgaccattg attatatctt tgttagcccg aaaaaccttg aagatgaagt tcaggatacc 240
catccggcat ttctggttcg taccctgatt acccgtagcc tgccgctgat tcatgatgaa 300
gttaaaaaac tgatcagcaa aagccgtctg attgccatta tttccgatgg tattattacc 360
caggttctgg aactggtgaa agatctgaat gttctgagct atacctattt tccgagcagc 420
gcaatgctgc tggcactgtg tctgtatagc gaaaatctgg atgaaaccac cacgagcgaa 480
tataaagatc tgctggaacc gatcaaaatt ccgggttgta ttccggttca gggtagcgat 540
ctgccggatc cgtttaatga tcgtaccagc gaaacctata aagaatttct ggaaggtagc 600
cgtcgttttt ttctggcaga tggtattctg gtgaacacct tttttgatct ggaagccagc 660
accattaaag aactgcaaga acaagaacgt cgtggtattg tgccgagcat tcatgcaatt 720
ggtccgtttg ttcagcatga aagcagcatg attgaaggca atgataataa caccctggaa 780
tgtctgaatt ggctggataa acagcaagaa aatagcgttc tgtatgtgag ctttggtagc 840
ggtggcacca ttagccataa acaaattatt gaactggccc tgggtttaga actgagcggt 900
cagaaattcc tgtggctgct gaaaccgcct agcaaatttg atatcatctt tgattttggc 960
cacttcagcg aagatccgct gaaatatctg ccgagcggtt ttctggaacg taccaaagaa 1020
cagggtatta ttgttccgta ttgggcaccg cagattaaaa tcctgggtca tgcagcaatt 1080
ggtggttatc tgtgtcattg tggttggaat agtattctgg aaagcgttgc acatggtatt 1140
ccgatgattg catggcctct gtttgcagaa cagcgtatga atgcagcact gttttgtaat 1200
ggtctgaaag ttgcaattcg tgccaaagtg aatgaaatgg gtattgttga acgtggtgaa 1260
gttgcgaaag tgatcaaaaa tctgatgatt ggtgatgaag gcaaagaaat tcgtcagcgt 1320
atgcgtgaac tgaaaggtag tgccgaagat gcaattaatg aaggtggtag cagcacccgt 1380
acactgaccc agctggtgca gaaatggaaa aacctggaat aa 1422
<210> 175
<211> 476
<212> PRT
<213> 胡麻(S. indicum)
<400> 175
Met Ser Ala Asp Gln Lys Leu Thr Ser Leu Val Phe Val Pro Phe Pro
1 5 10 15
Ile Met Ser His Leu Ala Thr Ala Val Lys Thr Ala Lys Leu Leu Ala
20 25 30
Asp Arg Asp Glu Arg Leu Ser Ile Thr Val Leu Val Met Lys Leu Pro
35 40 45
Ile Asp Thr Leu Ile Ser Ser Tyr Thr Lys Asn Ser Pro Asp Ala Arg
50 55 60
Val Lys Val Val Gln Leu Pro Glu Asp Glu Pro Thr Phe Thr Lys Leu
65 70 75 80
Met Lys Ser Ser Lys Asn Phe Phe Phe Arg Tyr Ile Glu Ser Gln Lys
85 90 95
Gly Thr Val Arg Asp Ala Val Ala Glu Ile Met Lys Ser Ser Arg Ala
100 105 110
Cys Arg Ile Ala Gly Phe Val Ile Asp Met Phe Cys Thr Pro Met Ile
115 120 125
Asp Val Ala Asn Glu Leu Gly Val Pro Thr Tyr Met Phe Phe Ser Ser
130 135 140
Gly Ser Ala Thr Leu Gly Leu Met Phe His Leu Gln Ser Leu Arg Asp
145 150 155 160
Asp Asn Asn Val Asp Val Met Glu Tyr Lys Asn Ser Asp Ala Ala Ile
165 170 175
Ser Ile Pro Thr Tyr Val Asn Pro Val Pro Val Ala Val Trp Pro Ser
180 185 190
Pro Val Phe Glu Glu Asp Ser Gly Phe Leu Asp Phe Ala Lys Arg Phe
195 200 205
Arg Glu Thr Lys Gly Ile Ile Val Asn Thr Phe Leu Glu Phe Glu Thr
210 215 220
His Gln Ile Arg Ser Leu Ser Asp Asp Lys Lys Ile Pro Pro Val Tyr
225 230 235 240
Pro Val Gly Pro Ile Leu Gln Ala Asp Glu Asn Lys Ile Glu Gln Glu
245 250 255
Lys Glu Lys His Ala Glu Ile Met Arg Trp Leu Asp Lys Gln Pro Asp
260 265 270
Ser Ser Val Val Phe Leu Cys Phe Gly Thr His Gly Cys Leu Glu Gly
275 280 285
Asp Gln Val Lys Glu Ile Ala Val Ala Leu Glu Asn Ser Gly His Arg
290 295 300
Phe Leu Trp Ser Leu Arg Lys Pro Pro Pro Lys Glu Lys Val Glu Phe
305 310 315 320
Pro Gly Glu Tyr Glu Asn Ser Glu Glu Val Leu Pro Glu Gly Phe Leu
325 330 335
Gly Arg Thr Thr Asp Met Gly Lys Val Ile Gly Trp Ala Pro Gln Met
340 345 350
Ala Val Leu Ser His Pro Ala Val Gly Gly Phe Val Ser His Cys Gly
355 360 365
Trp Asn Ser Val Leu Glu Ser Val Trp Cys Gly Val Pro Met Ala Val
370 375 380
Trp Pro Leu Ser Ala Glu Gln Gln Ala Asn Ala Phe Leu Leu Val Lys
385 390 395 400
Glu Phe Glu Met Ala Val Glu Ile Lys Met Asp Tyr Lys Lys Asn Ala
405 410 415
Asn Val Ile Val Gly Thr Glu Thr Ile Glu Glu Ala Ile Arg Gln Leu
420 425 430
Met Asp Pro Glu Asn Glu Ile Arg Val Lys Val Arg Ala Leu Lys Glu
435 440 445
Lys Ser Arg Met Ala Leu Met Glu Gly Gly Ser Ser Tyr Asn Tyr Leu
450 455 460
Lys Arg Phe Val Glu Asn Val Val Asn Asn Ile Ser
465 470 475
<210> 176
<211> 1431
<212> DNA
<213> 胡麻
<400> 176
atgagcgcag atcagaaact gaccagcctg gtttttgttc cgtttccgat tatgagccat 60
ctggcaaccg cagttaaaac cgcaaaactg ctggcagatc gtgatgaacg tctgagcatt 120
accgttctgg ttatgaaact gccgattgat accctgatta gcagctatac caaaaattca 180
ccggatgcgc gtgttaaagt tgttcagctg ccggaagatg aaccgacctt taccaaactg 240
atgaaaagca gcaaaaactt cttcttccgc tatatcgaaa gccagaaagg caccgttcgt 300
gatgcagttg cagaaattat gaaaagctca cgtgcatgtc gtattgccgg ttttgttatt 360
gatatgtttt gcaccccgat gattgatgtt gcaaatgaac tgggtgttcc gacctatatg 420
ttttttagca gcggtagcgc aaccctgggt ctgatgtttc atctgcagag cctgcgtgat 480
gataataatg ttgatgtgat ggaatacaaa aacagcgacg cagcaattag cattccgaca 540
tatgttaatc cggttccggt tgcagtttgg ccgagtccgg tttttgaaga agatagcggt 600
tttctggatt ttgccaaacg ttttcgtgaa accaaaggca ttattgtgaa cacgtttctg 660
gaatttgaaa cccatcagat tcgtagcctg tccgatgata aaaagattcc gcctgtttat 720
ccggttggtc cgattctgca ggccgatgaa aacaaaattg aacaagagaa agaaaaacac 780
gccgaaatta tgcgttggct ggataaacaa ccggattcaa gcgttgtttt tctgtgtttt 840
ggcacccatg gttgtctgga aggtgatcag gttaaagaaa ttgcagttgc cctggaaaat 900
agcggtcatc gttttctttg gagtctgcgt aaaccgcctc ctaaagaaaa agttgaattt 960
ccgggtgaat atgagaacag cgaagaagtt ctgcctgaag gctttctggg tcgtaccacc 1020
gatatgggta aagttattgg ttgggcaccg cagatggcag ttctgagtca tccggcagtt 1080
ggtggttttg tgagccattg tggttggaat agcgttctgg aaagcgtttg gtgtggtgtg 1140
ccgatggccg tttggcctct gagtgcagaa cagcaggcca atgcatttct gctggtgaaa 1200
gaattcgaaa tggccgtgga aatcaaaatg gactataaaa agaacgccaa cgttatcgtt 1260
ggtacggaaa ccattgaaga agcaattcgt cagctgatgg atccggaaaa tgaaattcgt 1320
gtgaaagttc gtgccctgaa agaaaagtca cgtatggcac tgatggaagg tggtagctca 1380
tataactatc tgaaacgctt tgtggaaaac gtggtgaaca acatcagcta a 1431
<210> 177
<211> 473
<212> PRT
<213> 葡萄(V. vinifera)
<400> 177
Met Glu Gln Thr Glu Leu Val Phe Ile Pro Phe Pro Val Ile Gly His
1 5 10 15
Leu Ala Ser Ala Leu Glu Ile Ala Lys Leu Ile Thr Lys Arg Asp Pro
20 25 30
Arg Phe Ser Ile Thr Ile Phe Ile Met Lys Phe Pro Phe Gly Ser Thr
35 40 45
Asp Gly Met Asp Thr Asp Ser Asp Ser Ile Arg Phe Val Thr Leu Pro
50 55 60
Pro Val Glu Val Ser Ser Glu Thr Thr Pro Ser Gly His Phe Phe Ser
65 70 75 80
Glu Phe Leu Lys Val His Ile Pro Leu Val Arg Asp Ala Val His Glu
85 90 95
Leu Thr Arg Ser Asn Ser Val Arg Leu Ser Gly Phe Val Ile Asp Met
100 105 110
Phe Cys Thr His Met Ile Asp Val Ala Asp Glu Phe Gly Val Pro Ser
115 120 125
Tyr Leu Phe Phe Ser Ser Gly Ala Ala Val Leu Gly Phe Leu Leu His
130 135 140
Val Gln Phe Leu His Asp Tyr Glu Gly Leu Asp Ile Asn Glu Phe Lys
145 150 155 160
Asp Ser Asp Ala Glu Leu Asp Val Pro Thr Phe Val Asn Ser Ile Pro
165 170 175
Gly Lys Val Phe Pro Ala Gly Met Phe Asp Lys Glu Ser Gly Gly Ala
180 185 190
Glu Met Leu Leu Tyr His Thr Arg Arg Phe Arg Glu Val Lys Gly Ile
195 200 205
Leu Val Asn Thr Phe Ile Glu Leu Glu Ser His Ala Ile Gln Ser Leu
210 215 220
Ser Gly Ser Thr Val Pro Glu Val Tyr Pro Val Gly Pro Ile Leu Asn
225 230 235 240
Thr Arg Met Gly Ser Gly Gly Gly Gln Gln Asp Ala Ser Ala Ile Met
245 250 255
Asn Trp Leu Asp Asp Gln Pro Pro Ser Ser Val Val Phe Leu Cys Phe
260 265 270
Gly Ser Met Gly Ser Phe Gly Ala Asp Gln Ile Lys Glu Ile Ala His
275 280 285
Ala Leu Glu His Ser Gly His Arg Phe Leu Trp Ser Leu Arg Gln Pro
290 295 300
Pro Pro Lys Gly Lys Met Ile Pro Ser Asp His Glu Asn Ile Glu Gln
305 310 315 320
Val Leu Pro Glu Gly Phe Leu His Arg Thr Ala Arg Ile Gly Lys Val
325 330 335
Ile Gly Trp Ala Pro Gln Ile Ala Val Leu Ala His Ser Ala Val Gly
340 345 350
Gly Phe Val Ser His Cys Gly Trp Asn Ser Leu Leu Glu Ser Val Trp
355 360 365
Tyr Gly Val Pro Val Ala Thr Trp Pro Ile Tyr Ala Glu Gln Gln Ile
370 375 380
Asn Ala Phe Gln Met Val Lys Asp Leu Gly Leu Ala Val Glu Ile Lys
385 390 395 400
Ile Asp Tyr Asn Lys Asp Arg Asp His Ile Val Ser Ala His Glu Ile
405 410 415
Glu Asn Gly Leu Arg Asn Leu Met Asn Ile Asn Ser Glu Val Arg Lys
420 425 430
Lys Arg Lys Glu Met Glu Lys Ile Ser His Lys Val Met Ile Asp Gly
435 440 445
Gly Ser Ser His Phe Ser Leu Gly His Phe Ile Glu Asp Met Asp Ser
450 455 460
Lys Val Met Lys Gly Lys Asp Ala Leu
465 470
<210> 178
<211> 1422
<212> DNA
<213> 葡萄
<400> 178
atggaacaga ccgaactggt gtttattccg tttccggtta ttggtcatct ggcaagcgca 60
ctggaaattg caaaactgat taccaaacgt gatccgcgtt ttagcattac catcttcatt 120
atgaaatttc cgtttggtag caccgatggt atggataccg atagcgatag cattcgtttt 180
gttaccctgc ctccggttga agttagcagc gaaaccacac cgagcggtca cttttttagc 240
gaatttctga aagttcatat tccgctggtt cgtgatgcag tgcatgaact gacccgtagc 300
aatagcgttc gtctgagcgg ttttgttatt gatatgtttt gcacccacat gattgatgtg 360
gcagatgaat ttggtgttcc gagctacctg ttttttagca gcggtgcagc agttctgggt 420
tttctgctgc atgttcagtt tctgcatgat tatgaaggcc tggatatcaa cgagtttaaa 480
gatagtgatg cggaactgga tgttccgacc tttgttaata gcattccggg taaagttttt 540
ccggcaggca tgtttgataa agaaagcggt ggtgcagaaa tgctgctgta tcacacccgt 600
cgttttcgtg aagttaaagg tattctggtg aacaccttta tcgaactgga aagccatgca 660
attcagagcc tgagcggtag taccgttccg gaagtttatc cggttggtcc gattctgaat 720
acccgtatgg gtagtggtgg tggtcagcag gatgcaagcg caattatgaa ttggctggat 780
gatcagcctc cgagcagcgt tgtttttctg tgttttggtt caatgggtag ctttggtgca 840
gatcagatta aagaaattgc acatgcactg gaacatagcg gtcatcgttt tctttggagc 900
ctgcgtcagc ctcctccgaa aggtaaaatg attccgagcg atcatgaaaa cattgaacag 960
gttctgccgg aaggctttct gcatcgtacc gcacgtattg gtaaagttat tggttgggca 1020
ccgcagattg ccgttctggc acatagcgca gttggtggtt ttgtgagcca ttgtggttgg 1080
aatagcctgc tggaaagcgt ttggtatggt gtgccggttg ccacctggcc gatttatgca 1140
gaacagcaga ttaatgcatt ccagatggtg aaagatctgg gtttagcagt ggaaatcaaa 1200
atcgactata acaaagatcg cgaccatatt gttagcgcac atgaaatcga aaatggtctg 1260
cgtaatctga tgaacattaa tagcgaagtg cgcaaaaaac gcaaagaaat ggaaaaaatc 1320
agccacaagg ttatgatcga tggtggtagc agccatttta gcctgggtca ttttattgaa 1380
gatatggaca gcaaagtgat gaaaggcaaa gatgcactgt aa 1422
<210> 179
<211> 470
<212> PRT
<213> 向日葵
<400> 179
Met Glu Arg Thr Pro His Ile Ala Ile Val Pro Ser Pro Gly Met Gly
1 5 10 15
His Leu Ile Pro Leu Val Glu Phe Ala Lys Arg Leu Lys Asn Asn His
20 25 30
Asn Ile Ser Ser Thr Phe Ile Ile Pro Asn Glu Gly Pro Leu Thr Lys
35 40 45
Ser Gln Gln Ala Phe Leu Asp Ser Leu Pro Asn Gly Leu Asn His Val
50 55 60
Ile Leu Pro Pro Val Ser Phe Asp Asp Leu Pro Asn Asp Ile Arg Met
65 70 75 80
Glu Thr Arg Ile Ser Leu Met Val Thr Arg Ser Leu Asp Ser Leu Arg
85 90 95
Glu Ala Val Lys Ser Leu Val Val Glu Thr Asn Met Val Ala Leu Phe
100 105 110
Val Asp Leu Phe Gly Thr Asp Ala Phe Asp Val Ala Ile Glu Phe Gly
115 120 125
Val Ser Pro Tyr Val Phe Phe Pro Ser Thr Ala Met Ala Leu Ser Leu
130 135 140
Phe Leu Tyr Leu Pro Lys Leu Asp Gln Met Val Ser Cys Glu Tyr Arg
145 150 155 160
Asp Leu Pro Glu Pro Val Gln Ile Pro Gly Cys Ile Pro Val Arg Gly
165 170 175
Glu Asp Leu Leu Asp Pro Val Gln Glu Arg Lys Asn Asp Ala Tyr Lys
180 185 190
Trp Val Leu His Asn Ala Lys Arg Tyr Arg Met Ala Glu Gly Ile Ala
195 200 205
Val Asn Ser Phe Lys Glu Leu Glu Gly Gly Ala Leu Lys Ala Leu Leu
210 215 220
Glu Asp Gln Pro Gly Lys Pro Arg Val Tyr Pro Val Gly Pro Leu Val
225 230 235 240
Gln Ala Gly Ser Ser Ser Asp Val Asp Gly Ser Gly Cys Leu Arg Trp
245 250 255
Leu Asp Gly Gln Pro Cys Gly Ser Val Leu Tyr Ile Ser Phe Gly Ser
260 265 270
Gly Gly Thr Leu Ser Ser Asn Gln Leu Asn Glu Leu Ala Leu Gly Leu
275 280 285
Glu Leu Ser Glu Gln Arg Phe Ile Trp Val Val Arg Ser Pro Asn Asp
290 295 300
Lys Pro Asn Ala Thr Tyr Phe Asn Ser His Gly His Glu Asp Pro Leu
305 310 315 320
Gly Phe Leu Pro Lys Gly Phe Leu Glu Arg Thr Lys Gly Ile Gly Phe
325 330 335
Val Val Pro Ser Trp Ala Pro Gln Ala Gln Ile Leu Ser His Ser Ser
340 345 350
Thr Gly Gly Phe Leu Thr His Cys Gly Trp Asn Ser Ile Leu Glu Thr
355 360 365
Val Val His Gly Val Pro Val Ile Ala Trp Pro Leu Tyr Ala Glu Gln
370 375 380
Arg Met Asn Ala Val Ser Leu Thr Glu Gly Ile Lys Val Ala Leu Arg
385 390 395 400
Pro Lys Val Asp Glu Asn Gly Ile Val Ser Arg Val Glu Ile Ala Arg
405 410 415
Val Val Lys Gly Leu Ile Glu Gly Glu Glu Gly Lys Pro Ile Arg Ser
420 425 430
Arg Ile Arg Glu Leu Lys Asp Ala Ala Ser Asn Val Leu Ser Lys Asp
435 440 445
Gly Cys Ser Thr Lys Thr Leu Glu Gln Leu Ala Ser Lys Leu Lys Ala
450 455 460
Lys Asn Asn Ile Ser Ile
465 470
<210> 180
<211> 1413
<212> DNA
<213> 向日葵
<400> 180
atggaacgta caccgcatat tgcaattgtt ccgagtcctg gtatgggtca tctgattccg 60
ctggttgaat ttgcaaaacg cctgaaaaac aaccacaata ttagcagcac ctttatcatt 120
ccgaatgaag gtccgctgac caaaagccag caggcatttc tggatagcct gccgaatggt 180
ctgaatcatg ttattctgcc tccggttagc tttgatgatc tgccgaacga tattcgtatg 240
gaaacccgta ttagcctgat ggttacccgt agcctggata gtctgcgtga agcagttaaa 300
agcctggttg ttgaaaccaa tatggttgca ctgtttgttg acctgtttgg caccgatgca 360
tttgatgttg caattgaatt tggtgttagc ccgtatgttt tttttccgag caccgcaatg 420
gcactgagcc tgtttctgta tctgcctaaa ctggatcaga tggttagctg tgaatatcgc 480
gatctgccgg aaccggtgca gattccgggt tgtattccgg ttcgtggtga agatctgctg 540
gatccggttc aagaacgtaa aaatgatgcc tataaatggg tgctgcataa cgcaaaacgt 600
tatcgtatgg cagaaggtat tgccgtcaat agctttaaag aactggaagg tggtgcactg 660
aaagcactgc tggaagatca gcctggtaaa ccgcgtgttt atccggttgg tccgctggtg 720
caggcaggta gcagcagtga tgttgatggt agcggttgtc tgcgttggct ggatggtcag 780
ccgtgtggta gcgttctgta tattagcttt ggtagtggtg gcaccctgag cagcaatcag 840
ctgaatgaac tggcactggg tttagaactg agcgaacagc gttttatttg ggttgttcgt 900
agccctaatg ataaaccgaa tgccacctat tttaacagcc atggtcatga agatcctctg 960
ggttttctgc cgaaaggttt tctggaacgc accaaaggta ttggttttgt tgtgccgagc 1020
tgggcaccgc aggcacagat tctgagccat agcagtaccg gtggttttct gacccattgt 1080
ggctggaata gcattctgga aaccgttgtt catggtgttc cggttattgc atggcctctg 1140
tatgcagaac agcgtatgaa tgcagttagc ctgaccgaag gtattaaagt tgcactgcgt 1200
ccgaaagttg atgaaaatgg tattgttagt cgtgtggaaa ttgcccgtgt tgttaaaggt 1260
ctgattgaag gtgaagaagg taaaccgatt cgtagccgta ttcgtgaact gaaagatgca 1320
gcaagcaatg ttctgagcaa agatggttgt agcaccaaaa cactggaaca gctggcaagc 1380
aaactgaaag ccaaaaacaa catcagcatt taa 1413
<210> 181
<211> 476
<212> PRT
<213> 潘那利番茄
<400> 181
Met Ser Pro Leu His Phe Phe Phe Phe Pro Met Val Ala Gln Gly His
1 5 10 15
Met Ile Pro Thr Leu Asp Met Ala Lys Leu Val Ala Ser Arg Gly Val
20 25 30
Lys Ala Thr Ile Ile Thr Thr Pro Leu Asn Glu Ser Val Phe Ser Asp
35 40 45
Ser Ile Glu Arg Asn Lys His Leu Gly Ile Glu Ile Asp Ile Arg Leu
50 55 60
Ile Thr Phe Gln Ala Val Glu Asn Asp Leu Pro Ile Gly Cys Glu Arg
65 70 75 80
Leu Asp Leu Val Pro Ser Pro Val Leu Phe Asn Asn Phe Phe Lys Ala
85 90 95
Thr Ala Met Met Gln Glu Pro Phe Glu Asn Leu Val Lys Glu Cys Arg
100 105 110
Pro Asp Cys Ile Val Ser Asp Met Leu Tyr Pro Trp Ser Thr Asp Ser
115 120 125
Ala Ala Lys Phe Asn Ile Pro Arg Ile Val Phe His Gly Thr Gly Phe
130 135 140
Phe Ala Leu Cys Val Ala Glu Ser Ile Lys Arg Asn Lys Pro Phe Lys
145 150 155 160
Asn Val Ser Thr Asp Ser Glu Thr Phe Val Val Pro Asn Leu Pro His
165 170 175
Gln Ile Arg Leu Thr Arg Thr Gln Leu Ser Pro Phe Asp Leu Glu Glu
180 185 190
Lys Glu Ala Ile Ile Phe Lys Ile Phe His Glu Val Arg Glu Ala Asp
195 200 205
Ser Lys Ser Tyr Gly Val Ile Phe Asn Ser Phe Tyr Glu Leu Glu Thr
210 215 220
Asp Tyr Phe Glu Tyr Tyr Thr Lys Phe Gln Asp Asn Lys Ser Trp Ala
225 230 235 240
Ile Gly Pro Leu Ser Leu Cys Asn Arg Tyr Ile Glu Asp Lys Ala Glu
245 250 255
Arg Gly Met Lys Ser Cys Ile Asp Thr His Glu Cys Leu Lys Trp Leu
260 265 270
Asp Ser Lys Lys Ser Gly Ser Ile Val Tyr Ile Cys Phe Gly Ser Gly
275 280 285
Val Thr Phe Thr Gly Ser Gln Ile Glu Glu Leu Ala Met Gly Ile Glu
290 295 300
Asp Ser Gly Gln Glu Phe Ile Trp Val Ile Arg Glu Gln Glu Asn Glu
305 310 315 320
Asn Ser Cys Leu Pro Glu Gly Phe Glu Glu Arg Thr Lys Glu Lys Gly
325 330 335
Leu Ile Ile Arg Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Glu
340 345 350
Gly Val Gly Ala Phe Val Thr His Cys Gly Trp Asn Ser Thr Leu Glu
355 360 365
Gly Ile Ser Ala Gly Val Pro Leu Val Ala Trp Pro Val Phe Ala Glu
370 375 380
Gln Phe Leu Asn Glu Lys Leu Val Thr Asp Val Leu Arg Ile Gly Val
385 390 395 400
Gly Val Gly Ser Val Lys Trp Glu Ala Ala Ala Ser Glu Gly Val Lys
405 410 415
Arg Glu Glu Ile Ser Lys Ala Ile Lys Arg Val Met Val Gly Glu Glu
420 425 430
Ala Glu Gly Phe Lys Asn Arg Ala Lys Glu Tyr Lys Glu Lys Ala Arg
435 440 445
Glu Ala Ile Glu Glu Gly Gly Ser Ser Tyr Asn Gly Leu Thr Asn Leu
450 455 460
Leu Gln Asp Val Ser Met Phe Gly Thr Lys Ile Asp
465 470 475
<210> 182
<211> 1431
<212> DNA
<213> 潘那利番茄
<400> 182
atgagtccgc tgcacttttt tttctttccg atggttgcac agggtcatat gattccgaca 60
ctggatatgg caaaactggt tgcaagccgt ggtgttaaag caaccattat taccacaccg 120
ctgaatgaaa gcgtttttag cgatagcatt gaacgcaata aacatctggg catcgaaatt 180
gatattcgcc tgattacctt tcaggccgtt gaaaatgatc tgccgattgg ttgtgaacgt 240
ctggatctgg ttccgagtcc ggttctgttt aataactttt tcaaagcaac cgccatgatg 300
caagaaccgt ttgaaaatct ggttaaagaa tgtcgtccgg attgcattgt tagcgatatg 360
ctgtatccgt ggtcaaccga tagcgcagcc aaatttaaca ttccgcgtat tgtttttcat 420
ggcaccggtt tttttgcact gtgtgttgca gaaagcatca aacgtaataa accgttcaaa 480
aacgttagca cggatagcga aacctttgtt gttccgaatc tgccgcatca gattcgtctg 540
acccgtacac agctgagccc gtttgatctg gaagaaaaag aagccatcat cttcaaaatc 600
tttcacgaag tgcgtgaagc agatagcaaa agctatggtg ttatcttcaa cagcttctat 660
gaactggaaa ccgactattt cgagtactac accaaattcc aggataacaa aagctgggca 720
attggtccgc tgagcctgtg taatcgttat atcgaagata aagcagagcg tggtatgaaa 780
agctgtattg atacccatga atgtctgaaa tggctggaca gcaaaaaatc aggtagcatt 840
gtgtatattt gctttggtag cggtgttacc tttaccggta gccagattga agaactggca 900
atgggtattg aagatagcgg tcaagaattt atctgggtga ttcgcgaaca agaaaatgaa 960
aatagctgtc tgccggaagg ttttgaagaa cgtaccaaag aaaaaggcct gattattcgt 1020
ggttgggcac cgcaggttct gattctggat catgaaggtg ttggtgcatt tgttacccat 1080
tgtggttgga atagcaccct ggaaggtatt agtgccggtg ttccgctggt tgcctggcct 1140
gtttttgcag aacagtttct gaacgaaaaa ctggtgaccg atgttctgcg tattggtgtt 1200
ggcgttggta gcgttaaatg ggaagcagca gcaagcgaag gtgttaaacg tgaagaaatt 1260
tccaaagcca ttaaacgtgt tatggttggt gaagaagccg aaggctttaa aaaccgtgcg 1320
aaagagtata aagagaaagc acgcgaagca attgaagaag gtggtagcag ctataatggt 1380
ctgaccaatc tgctgcagga tgttagcatg tttggcacca aaatcgatta a 1431
<210> 183
<211> 494
<212> PRT
<213> 糖甜菜(B. vulgaris)
<400> 183
Met Gly Ala Glu Pro Gln Arg Leu His Val Val Phe Phe Pro Leu Met
1 5 10 15
Ala Ala Gly His Leu Ile Pro Thr Leu Asp Ile Ala Lys Leu Phe Ala
20 25 30
Ala His His Val Lys Thr Thr Ile Ile Thr Thr Pro Leu Asn Ala Pro
35 40 45
Cys Phe Thr Lys Pro Leu Glu Ser Tyr Lys Asn Leu Gly His Arg Ile
50 55 60
Asp Ile Glu Ile Ile Pro Phe Pro Ser Lys Glu Ala Gly Leu Pro Glu
65 70 75 80
Gly Leu Glu Asn Phe Asp Gln Phe Thr Ser Asp Gln Met Ala Val Lys
85 90 95
Phe Leu Lys Ala Thr Glu Leu Leu Gln Glu Ser Phe Glu Lys Phe Leu
100 105 110
Glu Lys His Lys Pro Asn Cys Ile Val Thr Asp Met Leu Met Pro Phe
115 120 125
Thr Asn Asn Val Ala Ala Lys Phe Asn Ile Pro Arg Ile Val Phe His
130 135 140
Gly Cys Ser Tyr Phe Ala Leu Cys Met Met His Thr Leu Leu Lys Tyr
145 150 155 160
Gln Pro His Lys Ser Leu Leu Ser Asp Asp Glu Glu Phe Leu Val Pro
165 170 175
Asn Leu Pro His Glu Ile Asn Leu Thr Arg Ser Arg Leu Pro Asp Met
180 185 190
Met Arg Gly Gln Gly Asp Lys Glu Leu Asn Asp Ala Trp Met Lys Ile
195 200 205
Phe Ile His Ala Met Glu Ala Glu Glu Asn Ser Phe Gly Val Ile Met
210 215 220
Asn Ser Phe Tyr Glu Leu Glu Pro Glu Tyr Val Glu Tyr Tyr Arg Asn
225 230 235 240
Val Met Gly Arg Lys Ala Trp His Ile Gly Pro Val Ser Leu Cys Asn
245 250 255
Arg Glu Asn Glu Ala Lys Phe Gln Arg Gly Lys Asp Ser Ser Ile Asn
260 265 270
Glu His Glu Cys Leu Lys Trp Leu Asp Ser Lys Lys Pro Lys Ser Val
275 280 285
Val Tyr Ile Cys Phe Gly Ser Leu Ala Glu Val Pro Thr Leu Gln Leu
290 295 300
Arg Glu Ile Ala Met Gly Leu Glu Ala Ser Glu Gln Asp Phe Ile Trp
305 310 315 320
Val Val Arg Arg Gly Lys Glu Asn Val Glu Glu Glu Lys Ile Glu Glu
325 330 335
Trp Leu Pro Tyr Asp Phe Glu Asp Arg Met Glu Gly Lys Gly Leu Ile
340 345 350
Ile Arg Gly Trp Ala Pro Gln Val Leu Ile Leu Asp His Glu Ala Ile
355 360 365
Gly Ala Phe Val Thr His Cys Gly Trp Asn Ser Thr Leu Glu Gly Ile
370 375 380
Ser Cys Gly Val Pro Met Val Thr Trp Pro Val Phe Ala Glu Gln Phe
385 390 395 400
Tyr Asn Glu Lys Leu Val Thr Glu Val Leu Lys Thr Gly Val Ala Val
405 410 415
Gly Ala Lys Lys Trp Ser Arg Ile Leu Glu Val Asn Leu Lys Ser Glu
420 425 430
Asp Ile Lys Asn Ala Ile Arg Arg Val Met Val Gly Glu Glu Ala Leu
435 440 445
Val Leu Arg Ser Lys Ala Lys Lys Leu Lys Glu Leu Ala Arg Lys Ala
450 455 460
Val Glu Ile Gly Gly Ser Ser Tyr Ser Asp Met His Ser Leu Ile Gln
465 470 475 480
Asp Leu Ser Ser Tyr Asn Ala Asn Gly Tyr Lys Gln Tyr Leu
485 490
<210> 184
<211> 1485
<212> DNA
<213> 糖甜菜
<400> 184
atgggtgcag aaccgcagcg tctgcatgtt gttttttttc cgctgatggc agcaggtcat 60
ctgattccga cactggatat tgcaaaactg tttgcagcac atcatgtgaa aaccaccatt 120
attaccacac cgctgaatgc accgtgtttt acaaaaccgc tggaaagcta taaaaacctg 180
ggtcatcgta ttgacattga aattattccg tttccgagca aagaagcagg tctgccggaa 240
ggtctggaaa attttgatca gtttaccagc gatcagatgg ccgtgaaatt tctgaaagca 300
accgaactgc tgcaagaaag ctttgaaaaa ttcctggaaa aacacaagcc gaactgcatt 360
gttaccgata tgctgatgcc gtttaccaat aatgttgcag ccaaatttaa catccctcgc 420
attgtttttc atggctgtag ctattttgca ctgtgtatga tgcataccct gctgaaatat 480
cagccgcata aaagcctgct gagtgatgat gaagaatttc tggttccgaa tctgccgcat 540
gaaattaatc tgacccgtag tcgcctgccg gacatgatgc gtggtcaggg tgataaagaa 600
ctgaatgatg catggatgaa aatctttatc cacgcaatgg aagccgaaga aaatagcttt 660
ggtgtgatca tgaacagctt ctatgaactg gaaccggaat atgtggaata ctatcgtaat 720
gtgatgggtc gtaaagcatg gcatattggt ccggttagcc tgtgtaatcg tgaaaatgaa 780
gcaaaatttc agcgtggcaa agatagcagc attaacgaac atgaatgtct gaaatggctg 840
gacagcaaaa aaccgaaaag cgttgtgtat atttgctttg gtagcctggc agaagtgccg 900
acactgcagc tgcgtgaaat tgcaatgggt ttagaagcaa gcgaacagga tttcatttgg 960
gttgttcgtc gtggtaaaga aaacgtggaa gaagaaaaaa tcgaagagtg gctgccgtat 1020
gattttgaag atcgtatgga aggtaaaggc ctgattattc gtggttgggc accgcaggtt 1080
ctgattctgg atcatgaagc aattggtgca tttgttaccc attgtggttg gaatagcacc 1140
ctggaaggta ttagctgtgg tgttccgatg gttacctggc ctgtttttgc agaacagttc 1200
tataatgaaa aactggtgac cgaagttctg aaaaccggtg ttgcagttgg tgcaaaaaaa 1260
tggtcacgta ttctggaagt gaacctgaaa agcgaggata tcaaaaatgc aattcgtcgt 1320
gttatggttg gtgaagaagc actggttctg cgtagcaaag caaaaaaact gaaagaactg 1380
gcacgtaaag ccgttgaaat tggtggtagc agctatagcg atatgcatag cctgattcag 1440
gatctgagca gttataatgc caatggctat aaacagtatc tgtaa 1485
<210> 185
<211> 478
<212> PRT
<213> 毛果杨
<400> 185
Met Ala Glu Thr Asp Ser Pro Pro His Val Ala Ile Leu Pro Ser Pro
1 5 10 15
Gly Met Gly His Leu Ile Pro Leu Val Glu Leu Ala Lys Arg Leu Val
20 25 30
His Gln His Asn Leu Ser Val Thr Phe Ile Ile Pro Thr Asp Gly Ser
35 40 45
Pro Ser Lys Ala Gln Arg Ser Val Leu Gly Ser Leu Pro Ser Thr Ile
50 55 60
His Ser Val Phe Leu Pro Pro Val Asn Leu Ser Asp Leu Pro Glu Asp
65 70 75 80
Val Lys Ile Glu Thr Leu Ile Ser Leu Thr Val Ala Arg Ser Leu Pro
85 90 95
Ser Leu Arg Asp Val Leu Ser Ser Leu Val Ala Ser Gly Thr Arg Val
100 105 110
Val Ala Leu Val Val Asp Leu Phe Gly Thr Asp Ala Phe Asp Val Ala
115 120 125
Arg Glu Phe Lys Ala Ser Pro Tyr Ile Phe Tyr Pro Ala Pro Ala Met
130 135 140
Ala Leu Ser Leu Phe Phe Tyr Leu Pro Lys Leu Asp Glu Met Val Ser
145 150 155 160
Cys Glu Tyr Ser Glu Met Gln Glu Pro Val Glu Ile Pro Gly Cys Leu
165 170 175
Pro Ile His Gly Gly Glu Leu Leu Asp Pro Thr Arg Asp Arg Lys Asn
180 185 190
Asp Ala Tyr Lys Trp Leu Leu His His Ser Lys Arg Tyr Arg Leu Ala
195 200 205
Glu Gly Val Met Val Asn Ser Phe Ile Asp Leu Glu Arg Gly Ala Leu
210 215 220
Lys Ala Leu Gln Glu Val Glu Pro Gly Lys Pro Pro Val Tyr Pro Val
225 230 235 240
Gly Pro Leu Val Asn Met Asp Ser Asn Thr Ser Gly Val Glu Gly Ser
245 250 255
Glu Cys Leu Lys Trp Leu Asp Asp Gln Pro Leu Gly Ser Val Leu Phe
260 265 270
Val Ser Phe Gly Ser Gly Gly Thr Leu Ser Phe Asp Gln Ile Thr Glu
275 280 285
Leu Ala Leu Gly Leu Glu Met Ser Glu Gln Arg Phe Leu Trp Val Ala
290 295 300
Arg Val Pro Asn Asp Lys Val Ala Asn Ala Thr Tyr Phe Ser Val Asp
305 310 315 320
Asn His Lys Asp Pro Phe Asp Phe Leu Pro Lys Gly Phe Leu Asp Arg
325 330 335
Thr Lys Gly Arg Gly Leu Val Val Pro Ser Trp Ala Pro Gln Ala Gln
340 345 350
Val Leu Ser His Gly Ser Thr Gly Gly Phe Leu Thr His Cys Gly Trp
355 360 365
Asn Ser Thr Leu Glu Ser Val Val Asn Ala Val Pro Leu Ile Val Trp
370 375 380
Pro Leu Tyr Ala Glu Gln Lys Met Asn Ala Trp Met Leu Thr Lys Asp
385 390 395 400
Val Glu Val Ala Leu Arg Pro Lys Ala Ser Glu Asn Gly Leu Ile Gly
405 410 415
Arg Glu Glu Ile Ala Asn Ile Val Arg Gly Leu Met Glu Gly Glu Glu
420 425 430
Gly Lys Arg Val Arg Asn Arg Met Lys Asp Leu Lys Asp Ala Ala Ala
435 440 445
Glu Val Leu Ser Glu Ala Gly Ser Ser Thr Lys Ala Leu Ser Glu Val
450 455 460
Ala Arg Lys Trp Lys Asn His Lys Cys Thr Gln Asp Cys Asn
465 470 475
<210> 186
<211> 1437
<212> DNA
<213> 毛果杨
<400> 186
atggcagaaa ccgatagtcc gcctcatgtt gcaattctgc cgagtcctgg tatgggtcat 60
ctgattccgc tggttgaact ggcaaaacgt ctggttcatc agcataatct gagcgtgacc 120
tttattatcc cgaccgatgg tagcccgagc aaagcacagc gtagcgttct gggtagcctg 180
ccgagcacca ttcatagcgt ttttctgcct ccggttaatc tgagtgatct gccggaagat 240
gttaaaattg aaaccctgat tagcctgacc gttgcacgtt cactgccgag cctgcgtgat 300
gttctgagca gcctggttgc aagcggcacc cgtgttgttg cactggttgt tgacctgttt 360
ggcaccgatg catttgatgt tgcacgtgaa tttaaagcaa gcccgtatat cttttatccg 420
gcaccggcaa tggcactgag cctgtttttc tatctgccga aactggatga aatggtgagc 480
tgtgaatata gcgaaatgca agaaccggtt gaaattccgg gttgtctgcc gattcatggt 540
ggtgaactgc tggatccgac acgtgatcgt aaaaatgatg catataaatg gctgctgcat 600
cacagcaaac gttatcgtct ggccgaaggt gttatggtga atagctttat tgatctggaa 660
cgtggtgcac tgaaagcact gcaagaagtt gaaccgggta aaccgcctgt ttatccggtt 720
ggtccgctgg tgaatatgga tagcaatacc agcggtgttg aaggtagcga atgtctgaaa 780
tggctggatg atcagccgct gggtagcgtg ctgtttgtta gctttggtag cggtggcacc 840
ctgagctttg atcagattac cgaactggca ctgggtttag aaatgagcga acagcgtttt 900
ctgtgggttg cccgtgttcc gaatgataaa gttgcaaatg caacctattt cagcgtggat 960
aatcacaaag atccgtttga ttttctgccg aagggttttc tggatcgtac caaaggtcgt 1020
ggtctggttg ttccgagctg ggcaccgcag gcacaggttc tgagccatgg tagcaccggt 1080
ggttttctga cccattgtgg ttggaatagc accctggaaa gcgttgttaa tgcagttccg 1140
ctgattgttt ggcctctgta tgcagaacag aaaatgaatg catggatgct gaccaaagat 1200
gttgaagttg cactgcgtcc gaaagcaagc gaaaatggtc tgattggtcg tgaagaaatt 1260
gccaatattg tgcgtggtct gatggaaggt gaagaaggta aacgcgttcg taatcgtatg 1320
aaagatctga aagatgcagc cgcagaagtt ctgagcgaag caggtagcag caccaaagca 1380
ctgagtgaag ttgcccgtaa atggaaaaac cataaatgta cccaggactg caactaa 1437
<210> 187
<211> 469
<212> PRT
<213> Q. suber
<400> 187
Met Glu Gln Lys Pro His Ile Ala Leu Leu Pro Ser Pro Gly Met Gly
1 5 10 15
His Leu Ile Pro Leu Val Glu Phe Ala Lys Gln Phe Val Leu His His
20 25 30
Asp Phe His Ile Thr Cys Ile Ile Pro Val Leu Gly Ser Pro Ser Lys
35 40 45
Ala Met Lys Ala Val Leu Gln Ala Leu Pro Thr Thr Ile Asp His Val
50 55 60
Phe Leu Pro Pro Val Ile Leu Glu Glu Glu Glu Ile Lys Gly Leu Lys
65 70 75 80
Phe Glu Val Gln Thr Ile Leu Thr Leu Thr Arg Ser Leu Pro Pro Leu
85 90 95
Arg Glu Val Leu Lys Thr Thr Arg Phe Ser Ala Phe Val Val Asp Pro
100 105 110
Phe Gly Ile Asp Ala Leu Asp Ile Ala Lys Glu Leu Asn Ile Ser Pro
115 120 125
Tyr Ile Phe Phe Pro Ser Asn Ala Phe Ala Leu Ser Leu Ile Phe His
130 135 140
Leu Pro Lys Leu Asp Glu Thr Val Ser Cys Glu Tyr Arg Asp Leu Pro
145 150 155 160
Glu Pro Leu Lys Leu Pro Gly Cys Ile Pro Ile His Gly Arg Asp Leu
165 170 175
Ile Glu Pro Val Gln Asp Arg Thr Ser Glu Leu Tyr Lys Met Phe Leu
180 185 190
Arg Asn Ala Lys Arg Phe Arg Leu Ala Glu Gly Ile Ile Val Asn Thr
195 200 205
Phe Met Glu Leu Glu Gly Ser Ala Ile Lys Ala Leu Leu Asp Glu Glu
210 215 220
Ala Lys Asn Leu Pro Leu Tyr Pro Ile Gly Pro Ile Gln Ser Gly Ser
225 230 235 240
Ser Asn Leu Gln Val Asp Lys Ser Val Ser Asp Cys Leu Arg Trp Leu
245 250 255
Asp Asn Gln Pro His Gly Ser Val Leu Phe Val Cys Phe Gly Ser Gly
260 265 270
Gly Thr Leu Ser Tyr Asp Gln Thr Asn Glu Leu Ala Leu Gly Leu Glu
275 280 285
Leu Ser Gly Gln Lys Phe Leu Trp Val Val Arg Thr Pro Asn Asn Glu
290 295 300
Ser Ala Asp Ala Ala Tyr Leu Ser Asp Gln Ile Leu Asp Asn Asn Pro
305 310 315 320
Leu Asp Phe Leu Pro Lys Gly Phe Val Glu Arg Thr Glu Gly Gln Gly
325 330 335
Leu Ala Val Pro Ser Trp Ala Pro Gln Ala Gln Val Leu Ser His Gly
340 345 350
Ser Thr Gly Gly Phe Leu Thr His Cys Gly Trp Asn Ser Thr Leu Glu
355 360 365
Ser Ile Met Gln Gly Ile Pro Leu Ile Ala Trp Pro Leu Tyr Ala Glu
370 375 380
Gln Lys Met Asn Ala Pro Leu Leu Ala Glu Asp Leu Lys Val Ala Leu
385 390 395 400
Arg Pro Lys Thr Asn Lys Ser Gly Leu Ile Asp Gln Glu Glu Ile Ala
405 410 415
Lys Val Val Lys Gly Leu Met Ile Gly Glu Glu Gly Lys Lys Val Tyr
420 425 430
Asn Arg Met Lys Asp Ile Lys Met Ala Ala Glu Lys Ala Leu Ser Ala
435 440 445
Asp Gly Ser Ser Thr Lys Ala Leu Ser Glu Leu Ala Ser Gln Trp Lys
450 455 460
Asn His Pro Gly Phe
465
<210> 188
<211> 1410
<212> DNA
<213> Q. suber
<400> 188
atggaacaga aaccgcatat tgcactgctg ccgagtcctg gtatgggtca tctgattccg 60
ctggttgaat ttgcaaaaca gtttgtgctg catcatgatt tccatatcac ctgtattatt 120
ccggttctgg gtagcccgag caaagcaatg aaagcagttc tgcaggcact gccgaccacc 180
attgatcatg tttttctgcc tccggttatt ctggaagaag aagaaattaa aggcctgaaa 240
tttgaagtgc agaccattct gaccctgaca cgtagcctgc ctccgctgcg tgaagttctg 300
aaaaccacac gttttagcgc atttgttgtt gatccgtttg gtattgatgc actggatatt 360
gccaaagaac tgaacattag cccgtatatc ttttttccga gcaatgcatt tgcactgagc 420
ctgatttttc atctgccgaa actggatgaa accgttagct gtgaatatcg tgatctgccg 480
gaaccgctga aactgcctgg ttgtattccg attcatggtc gcgatctgat tgaaccggtg 540
caggatcgta ccagcgaact gtataaaatg tttctgcgta atgccaaacg ttttcgtctg 600
gcagaaggca ttattgtcaa tacctttatg gaactggaag gcagcgcaat taaagcactg 660
ctggatgaag aagcaaaaaa tctgccgctg tatccgattg gtccgattca gagcggtagc 720
agcaatctgc aggttgataa aagcgttagc gattgtctgc gttggctgga taatcagccg 780
catggtagcg ttctgtttgt ttgttttggt agcggtggca ccctgagcta tgatcagacc 840
aatgaactgg cactgggttt agaactgagc ggtcagaaat tcctgtgggt tgttcgtacc 900
ccgaataatg aaagcgcaga tgcagcatat ctgagcgatc agattctgga taataatccg 960
ctggattttc tgccaaaagg ttttgttgaa cgtaccgaag gtcaaggtct ggcagttccg 1020
agctgggcac cgcaggcaca ggttctgagc catggtagca ccggtggttt tctgacccat 1080
tgtggttgga atagcaccct ggaaagcatt atgcagggta ttccgctgat tgcatggcct 1140
ctgtatgcag aacagaaaat gaatgcaccg ctgctggccg aagatctgaa agttgcactg 1200
cgtccgaaaa ccaataaaag cggtctgatt gatcaagaag agatcgccaa agttgttaag 1260
ggtctgatga ttggtgaaga gggcaaaaaa gtgtacaatc gcatgaaaga cattaagatg 1320
gcagcagaaa aagcactgag tgcagatggt agcagtacca aagcgctgag cgaactggca 1380
agccagtgga aaaatcatcc gggtttttaa 1410
<210> 189
<211> 475
<212> PRT
<213> A. duranensis
<400> 189
Met Ala Lys Thr Met Arg Ile Ala Val Ile Thr Ser Pro Gly Leu Thr
1 5 10 15
His Leu Val Pro Ile Leu Glu Phe Ser Lys Arg Phe Leu Glu Leu His
20 25 30
Pro Asn Phe His Val Thr Cys Met Ile Pro Ser Leu Gly Pro His Pro
35 40 45
Asp Ser Thr Lys Ser Tyr Leu Gln Thr Leu Pro Ser Asn Ile His Ser
50 55 60
Ile Leu Leu Pro Pro Ile Asn Lys Gln Asp Leu Pro Gln Gly Ala Tyr
65 70 75 80
Pro Gly Val Leu Ile Gln Lys Thr Val Thr Leu Ser Leu Pro Ser Ile
85 90 95
Arg Asp Thr Leu Lys Ser Leu Thr Leu Arg Glu Pro Leu Ala Ala Leu
100 105 110
Ile Ala Asp Ala Tyr Ala Phe Glu Ala Leu Ser Phe Ala Lys Glu Phe
115 120 125
Asn Phe Leu Ser Tyr Ile Tyr Phe Pro Ser Ser Val Met Ala Leu Ser
130 135 140
Leu Cys Leu His Leu Pro Lys Leu Asp Glu Gln Val Thr Gly Glu Tyr
145 150 155 160
Lys Asp Leu Lys Asp Pro Ile Tyr Leu Pro Gly Cys Val Pro Val Phe
165 170 175
Gly Arg Asp Leu Pro Phe Pro Met Gln Asn Arg Ser Ser Asp Ala Tyr
180 185 190
Lys Leu Tyr Leu Glu Arg Ser Lys Gly Phe Ser Asn Val Asp Gly Phe
195 200 205
Ile Ile Asn Ser Phe Leu Glu Leu Glu Ser Ala Ala Met Lys Ala Leu
210 215 220
Ala Arg Glu Lys Ser Cys Phe Ser Phe Tyr Asp Val Gly Pro Ile Thr
225 230 235 240
Gln Lys Arg Ser Ser Ser Asn Asp Gly Asp Glu Glu Leu Glu Cys Leu
245 250 255
Arg Trp Leu Asp Lys Gln Pro His Ser Ser Val Leu Tyr Val Ser Phe
260 265 270
Gly Ser Gly Gly Thr Leu Ser Gln Ser Ala Ile Asn Glu Leu Ala Phe
275 280 285
Gly Leu Glu Leu Ser Gly Gln Arg Phe Leu Trp Val Leu Arg Ala Pro
290 295 300
Ser Asp Ser Ser Ser Ala Ala Tyr Leu Asp Asn Gln Lys Asn Glu Asp
305 310 315 320
Pro Leu Lys Phe Leu Pro Ser Gly Phe Leu Glu Arg Thr Lys Glu Lys
325 330 335
Gly Leu Val Leu Pro Ser Trp Ala Pro Gln Val Gln Ile Leu Ser His
340 345 350
Asp Ser Val Gly Gly Phe Leu Ser His Cys Gly Trp Asn Ser Val Leu
355 360 365
Glu Ser Val Gln Val Gly Val Pro Ile Ile Thr Trp Pro Leu Phe Ala
370 375 380
Glu Gln Arg Met Asn Ala Val Leu Leu Val Asp Gly Leu Lys Val Ala
385 390 395 400
Val Arg Pro Asn Val Gly Glu Asp Gly Val Val Gly Lys Glu Glu Val
405 410 415
Ser Asn Val Ile Lys Cys Leu Met Glu Gln Glu Glu Gly Lys Ala Met
420 425 430
Arg Lys Arg Met Glu Asp Leu Lys Ala Tyr Ala Ala Asp Ala Val Asn
435 440 445
Lys Asp Ala Gly Ser Ser Thr His Ala Leu Ser His Leu Ala Thr Lys
450 455 460
Trp Glu Asn Phe Ser Gly Ile Glu Asp Asn Asn
465 470 475
<210> 190
<211> 1428
<212> DNA
<213> A. duranensis
<400> 190
atggcaaaaa ccatgcgtat tgccgttatt accagtccgg gtctgaccca tctggttccg 60
attctggaat ttagcaaacg ttttctggaa ctgcatccga attttcatgt tacctgtatg 120
attccgagcc tgggtccgca tccggatagc accaaaagct atctgcagac cctgccgagc 180
aatattcata gcattctgct gcctccgatt aacaaacagg atctgccgca gggtgcatat 240
ccgggtgttc tgattcagaa aaccgttaca ctgagcctgc cgagtattcg tgataccctg 300
aaaagtctga ccctgcgtga accgctggca gcactgattg cagatgcata tgcctttgaa 360
gcactgagct ttgccaaaga attcaacttt ctgagctata tctatttccc gagcagcgtt 420
atggccctga gcctgtgtct gcatctgccg aaactggatg aacaggttac cggtgaatat 480
aaagatctga aagatccgat ttatctgcct ggttgtgttc cggtttttgg tcgtgatctg 540
ccgtttccga tgcagaatcg tagcagtgat gcatataaac tgtatctgga acgcagcaaa 600
ggttttagca atgtggatgg ctttatcatc aacagctttc ttgaactgga aagcgcagca 660
atgaaagcac tggcacgtga aaaaagctgc tttagctttt atgatgtggg tccgattaca 720
cagaaacgta gctcaagcaa tgatggtgat gaagaactgg aatgtctgcg ttggctggat 780
aaacagccgc atagcagcgt tctgtatgtt agctttggta gcggtggcac cctgagccag 840
agcgcaatta atgaactggc atttggcctg gaactgagcg gtcagcgttt tctgtgggtt 900
ctgcgtgcac cgagcgatag cagcagcgca gcatatctgg ataatcagaa aaatgaagat 960
ccgctgaaat ttctgccgag cggtttcctg gaacgtacca aagaaaaagg tctggtgctg 1020
ccgagctggg caccgcaggt tcagattctg agccatgata gcgttggtgg ttttctgtca 1080
cattgtggtt ggaatagcgt tctggaaagt gttcaggttg gtgttccgat tattacctgg 1140
cctctgtttg cagaacagcg tatgaatgca gttctgctgg ttgatggtct gaaagttgca 1200
gttcgtccga atgttggtga agatggtgtt gttggtaaag aagaagttag caacgttatc 1260
aagtgcctga tggaacaaga agagggtaaa gcaatgcgta aacgtatgga agatttaaaa 1320
gcatatgcag ccgatgccgt taataaagat gcaggtagca gcacccatgc actgagccat 1380
ctggcaacca aatgggaaaa ctttagcggt attgaggaca acaactaa 1428
<210> 191
<211> 495
<212> PRT
<213> 番木瓜
<400> 191
Met Gly Ser Glu Val Leu His His Asp Tyr Ser Gln Leu Asn Ile Phe
1 5 10 15
Phe Phe Pro Phe Met Ala His Gly His Met Ile Pro Thr Leu Asp Met
20 25 30
Ala Lys Leu Phe Ala Thr His Gly Ala Lys Thr Ser Ile Ile Thr Thr
35 40 45
Pro Leu Asn Leu Pro Phe Phe Ser Lys Ser Ile Glu Arg Phe Ser Lys
50 55 60
Gln Thr Gly Leu Glu Ile Gly Val Lys Leu Leu Asn Phe Pro Ser Val
65 70 75 80
Glu Val Gly Leu Pro Ser Gly Cys Glu Asn Ala Asp Ser Leu Pro Ala
85 90 95
Gly Glu Pro Leu Ile Val Asn Lys Phe Phe Ala Ala Ala Gly Met Leu
100 105 110
Lys Asp Pro Leu Glu Arg Leu Leu Gln Glu Phe Lys Pro Asp Cys Leu
115 120 125
Ile Ala Asp Met Phe Phe Pro Trp Thr Thr Asp Ala Ala Ala Lys Phe
130 135 140
Asp Ile Pro Arg Leu Val Phe His Gly Thr Ser Phe Phe Ala Leu Ser
145 150 155 160
Ala Ser Glu Cys Ile Arg Leu Tyr Thr Pro Phe Asn Asn Val Ser Ser
165 170 175
Asp Ser Glu Pro Phe Leu Val Pro Thr Leu Pro Asp Glu Ile Arg Leu
180 185 190
Thr Arg Asn Gln Leu Ala Asp Phe Ala Met Lys Glu Gly Asp Glu Asn
195 200 205
Gly Ile His Arg Leu Ile Lys Glu Ala Lys Glu Ser Glu Leu Lys Ser
210 215 220
Tyr Gly Val Val Val Asn Ser Phe Tyr Glu Leu Glu Pro Ala Tyr Ala
225 230 235 240
Asp His Tyr Arg Asn Phe Leu Lys Arg Lys Ala Trp His Ile Gly Pro
245 250 255
Val Ser Leu Cys Asn Lys Thr Val Glu Asp Lys Ala Glu Arg Gly Lys
260 265 270
Arg Ala Ser Ile Asp Glu Asp Glu Cys Leu Lys Trp Leu Asn Ser Lys
275 280 285
Ala Pro Asn Ser Val Ile Tyr Ile Cys Phe Gly Ser Met Ala Asn Phe
290 295 300
Asn Ser Ala Gln Leu Met Glu Ile Ala Thr Ala Leu Asp Ala Ser Gly
305 310 315 320
Gln Glu Phe Ile Trp Val Val Arg Arg Glu Lys Asn Glu Asn Asn Gln
325 330 335
Glu Asp Trp Leu Pro Glu Gly Phe Glu Gln Arg Thr Glu Gly Lys Gly
340 345 350
Leu Ile Ile Arg Gly Trp Ala Pro Gln Val Leu Ile Leu Glu His Glu
355 360 365
Ala Val Gly Gly Phe Val Thr His Cys Gly Trp Asn Ser Thr Leu Glu
370 375 380
Gly Val Thr Ala Gly Met Pro Met Val Thr Trp Pro Val Ser Ala Glu
385 390 395 400
Gln Phe Tyr Asn Glu Lys Leu Val Thr Glu Val Leu Lys Ile Gly Leu
405 410 415
Ser Val Gly Val Lys Lys Trp Val Arg Ser Glu Gly Asp Phe Val Ser
420 425 430
Arg Glu Lys Val Glu Gln Ala Val Arg Glu Ile Met Val Gly Ser Glu
435 440 445
Ala Val Glu Arg Arg Met Arg Ala Lys Ala Met Ala Asp Met Ala Arg
450 455 460
Ala Ala Val Glu Lys Gly Gly Ser Ser Tyr Asn Asp Leu Asn Ala Leu
465 470 475 480
Leu Arg Glu Val Ser Leu Met Arg Arg Gln Gln Ser Gln Asn Gln
485 490 495
<210> 192
<211> 1488
<212> DNA
<213> 番木瓜
<400> 192
atgggtagcg aagttctgca tcatgattat agccagctga acatcttttt ctttccgttt 60
atggcacatg gtcatatgat tccgacactg gatatggcaa aactgtttgc aacccatggt 120
gcaaaaacca gcattattac cacaccgctg aatctgccgt tttttagcaa aagcattgaa 180
cgctttagca aacagacagg tctggaaatt ggtgtgaaac tgctgaattt tccgagcgtt 240
gaagttggtc tgccgagcgg ttgtgaaaat gcagatagcc tgcctgccgg tgaaccgctg 300
attgtgaata aattctttgc agcagcaggc atgctgaaag atccgctgga acgtctgctg 360
caagagttta aaccggattg tctgattgcc gatatgtttt ttccgtggac caccgatgca 420
gcagccaaat ttgatattcc gcgtctggtt tttcatggca ccagcttttt tgcactgagc 480
gcaagcgaat gtattcgtct gtataccccg tttaataacg ttagcagcga tagcgaaccg 540
tttctggtgc cgacactgcc ggatgaaatt cgtctgaccc gtaatcagct ggcagatttt 600
gcaatgaaag aaggtgacga aaacggtatt catcgtctga ttaaagaagc caaagaaagc 660
gagctgaaaa gctatggtgt tgtggtgaat agcttttatg aactggaacc ggcatatgcg 720
gatcattatc gtaattttct gaaacgcaaa gcctggcata ttggtccggt tagcctgtgt 780
aataaaaccg ttgaagataa agccgaacgt ggtaaacgtg caagcattga tgaagatgaa 840
tgtctgaaat ggctgaatag caaagcaccg aatagcgtga tttatatctg ctttggtagc 900
atggccaatt ttaacagcgc acagctgatg gaaattgcaa ccgcactgga tgcaagcggt 960
caagaattca tttgggttgt tcgtcgcgaa aaaaacgaaa acaatcaaga agattggctg 1020
ccggaaggtt ttgaacagcg taccgaaggt aaaggtctga ttattcgtgg ttgggcaccg 1080
caggttctga ttctggaaca tgaagcagtt ggtggttttg ttacccattg tggttggaat 1140
agcaccctgg aaggtgttac cgcaggtatg ccgatggtta cctggcctgt tagcgcagaa 1200
cagttttata acgaaaaact ggttaccgag gtgctgaaaa ttggtctgag cgtgggtgtg 1260
aaaaaatggg ttcgtagcga aggtgatttt gtgagccgtg aaaaagttga acaggcagtt 1320
cgtgaaatta tggttggtag tgaagccgtt gaacgtcgta tgcgtgcaaa agcaatggca 1380
gatatggcac gtgcagcagt tgaaaaaggt ggtagcagct ataatgatct gaatgcactg 1440
ctgcgtgaag ttagcctgat gcgtcgtcag cagagtcaga atcagtaa 1488
<210> 193
<211> 491
<212> PRT
<213> Z. jujube
<400> 193
Met Lys Lys Ala Glu Leu Val Phe Ile Pro Ile Pro Gly Arg Gly His
1 5 10 15
Leu Leu Ser Met Val Glu Phe Ala Lys Leu Leu Val Ala Arg Asp Pro
20 25 30
His Leu Tyr Val Thr Ile Leu Ile Met Lys Leu Pro Phe Asp Thr Lys
35 40 45
Val Gly Ala Tyr Thr Ala Ser Leu Val Ser Ser Ser Ser Asn Arg Ile
50 55 60
Asn Cys Ile Asp Leu Pro Ile Asn Glu Lys Val Tyr Thr Glu Ser Asn
65 70 75 80
Pro Pro Val Phe Met Thr Ser Phe Ile Glu Asp Gln Lys Pro His Val
85 90 95
Lys Asn Ala Val Thr Gln Leu Ile Gln Ser Arg Asp Val Asp Asp Glu
100 105 110
Asp Ser Pro Arg Leu Ala Gly Phe Val Ile Asp Met Phe Cys Thr Thr
115 120 125
Met Ile Asp Val Ala Asn Glu Phe Gly Ile Pro Thr Tyr Val Phe Phe
130 135 140
Ala Ser Gly Ala Gly Phe Leu Gly Leu Leu Phe His Leu Gln His Leu
145 150 155 160
Ser Asp Asn His Asn Val Asn Ile Thr Glu Phe Glu Asn Asp Pro Glu
165 170 175
Ala Glu Leu Val Ile Pro Ser Phe Val Asn Pro Phe Pro Ser Lys Val
180 185 190
Leu Pro Val Leu Val Leu Asp Lys Asp Gly Gly Pro Val Met Met Asn
195 200 205
His Ala Arg Arg Ile Arg Glu Thr Lys Gly Ile Ile Val Asn Thr Phe
210 215 220
Ile Glu Leu Glu Ser His Ala Val Tyr Ser Leu Ser Asn Gly Asp His
225 230 235 240
Glu Phe Pro Pro Val Tyr Pro Val Gly Pro Ile Leu Tyr Leu Lys Ser
245 250 255
Asp Glu Ser His Val Gly Ser Val Asn Gln Ile Gln Asn Ser Asp Ile
260 265 270
Ile Arg Trp Leu Asp Asn Gln Pro Pro Ser Ser Val Val Phe Val Cys
275 280 285
Phe Gly Ser Met Gly Ser Phe Ser Glu Asp Gln Val Lys Glu Ile Ala
290 295 300
Tyr Gly Leu Glu Gln Ser Gly Gln Arg Phe Ile Trp Ser Leu Arg Pro
305 310 315 320
Pro Pro Pro Lys Asp Lys Met Gly Phe Pro Ser Asp Tyr Leu Asp Pro
325 330 335
Thr Val Val Leu Pro Glu Gly Phe Leu Asp Arg Thr Ala Glu Val Gly
340 345 350
Lys Val Ile Gly Trp Ala Pro Gln Val Glu Ile Leu Ser His Cys Ala
355 360 365
Thr Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Thr Leu Glu Ser
370 375 380
Leu Trp Phe Gly Val Pro Ile Ala Thr Trp Pro Ile Phe Ala Glu Gln
385 390 395 400
Gln Leu Asn Ala Phe Gln Met Val Lys Glu Phe Gly Cys Ala Val Glu
405 410 415
Ile Lys Leu Asp Tyr Arg Arg Glu Phe Asn Ser Asp Gly Asp Asp Gln
420 425 430
Ala Val Val Ser Ala Gln Glu Ile Glu Arg Gly Ile Arg Arg Val Met
435 440 445
Asp Asp Asp Ser Asp Ile Arg Lys Arg Thr Lys Glu Ile Ser Glu Gln
450 455 460
Ser Arg Arg Thr Leu Val Asp Gly Gly Thr Ser Phe Ser Cys Leu Gly
465 470 475 480
His Leu Ile Asn Asp Ile Leu Glu Asn Val Ser
485 490
<210> 194
<211> 1476
<212> DNA
<213> Z. jujube
<400> 194
atgaaaaaag ccgaactggt gtttattccg attcctggtc gtggtcatct gctgagcatg 60
gttgaatttg caaaactgct ggttgcacgt gatccgcatc tgtatgttac cattctgatt 120
atgaaactgc cgttcgatac caaagttggt gcatataccg caagcctggt tagcagcagc 180
agtaatcgta ttaattgtat tgatctgccg atcaacgaga aagtgtatac cgaaagcaat 240
ccgcctgttt ttatgaccag ctttatcgaa gatcagaaac cgcatgttaa aaatgcagtt 300
acccagctga ttcagagccg tgatgttgat gatgaagata gtccgcgtct ggcaggtttt 360
gttattgata tgttttgcac caccatgatc gatgtggcaa atgaatttgg tattccgacc 420
tatgtttttt ttgcaagcgg tgcaggtttt ctgggtctgc tgtttcatct gcagcatctg 480
agcgataatc ataacgtgaa catcaccgaa tttgagaatg atccggaagc agaactggtt 540
attccgagct ttgttaatcc gtttccgagc aaagttctgc cggttctggt tctggataaa 600
gatggtggtc cggttatgat gaatcatgca cgtcgtattc gtgaaaccaa aggcattatt 660
gtgaacacct ttattgaact ggaaagccat gcagtttata gcctgagcaa tggtgatcat 720
gaatttccgc cagtttatcc ggttggtccg attctgtatc tgaaaagtga tgaaagtcat 780
gtgggtagcg ttaatcagat tcagaacagc gatattattc gctggctgga taatcagcct 840
ccgagcagcg ttgtttttgt ttgttttggt agcatgggta gctttagtga ggatcaggtt 900
aaagaaattg cctatggtct ggaacagagc ggtcagcgtt ttatttggag cctgcgtccg 960
cctccgccta aagataaaat gggttttccg agcgattatc tggatccgac cgttgtgctg 1020
ccggaaggct ttctggatcg taccgcagaa gttggtaaag ttattggttg ggcaccgcag 1080
gttgaaattc tgagccattg tgcaaccggt ggttttgttt cacattgtgg ttggaatagc 1140
accctggaaa gtctgtggtt tggtgttccg attgcaacct ggccgatttt tgcagaacag 1200
cagctgaatg catttcagat ggtgaaagaa tttggttgtg ccgtggaaat caaactggat 1260
tatcgtcgtg aatttaacag cgacggtgat gatcaggcag ttgttagcgc acaagaaatt 1320
gaacgtggta ttcgtcgtgt tatggatgat gatagcgata ttcgtaaacg caccaaagaa 1380
attagcgaac agagccgtcg taccctggtt gatggtggta caagctttag ctgtctgggt 1440
catctgatca atgatattct ggaaaacgtg agctaa 1476
<210> 195
<211> 483
<212> PRT
<213> 向日葵
<400> 195
Met Ala Asn Ala Val Ala Glu Leu Ile Phe Ile Pro Thr Pro Gly Leu
1 5 10 15
Gly His Ile Met Ser Thr Ile Glu Leu Ala Lys Leu Leu Val Asn Arg
20 25 30
Asp Gln Arg Leu Ala Ile Thr Val Leu Val Ile Lys Pro Pro Gly Met
35 40 45
Thr Ser Gly Ser Ala Ile Thr Thr Tyr Ile Glu Ser Leu Thr Glu Thr
50 55 60
Thr Met Asp Arg Ile Ser Phe Ile Gln Leu Pro Gln Val Glu Ser Ser
65 70 75 80
Pro Thr His Gly Gly Pro Thr Glu Phe Ile Arg Ser His Ser Lys Tyr
85 90 95
Val Arg Asn Ala Val Val Asp Leu Arg Ser Gln Ser Gly Ser Cys Gln
100 105 110
Val Val Gly Phe Val Val Asp Met Phe Cys Thr Ser Met Ile Asp Val
115 120 125
Ala Asn Glu Phe Asn Val Pro Thr Phe Val Phe Phe Thr Ser Ser Ala
130 135 140
Ala Phe Leu Gly Phe Thr Leu Phe Ile Lys Leu Leu Cys Asp Asp Leu
145 150 155 160
Asn Arg Asp Val Val Glu Leu Ser Asn Ser Asp Thr Glu Ile Ser Val
165 170 175
Pro Ser Phe Val Lys Pro Val Pro Thr Lys Val Phe Trp Ser Leu Val
180 185 190
Lys Thr Arg Glu Gly Leu Asp Ser Val Gln Arg Leu Ala Lys Lys Leu
195 200 205
Gly Glu Ala Lys Gly Ile Ile Val Asn Thr Phe Leu Asp Leu Glu Thr
210 215 220
His Ala Ile Glu Ser Leu Ser Ala Asp Ile Ser Ile Pro Pro Val Tyr
225 230 235 240
Pro Val Gly Pro Ile Leu Asn Leu Glu Gly Gly Ser Gly Gly Gly Lys
245 250 255
Pro Phe Asp Asp Asp Val Ile Arg Trp Leu Asp Ser Gln Pro Pro Ser
260 265 270
Ser Val Val Phe Leu Cys Phe Gly Ser Met Gly Ser Phe Asp Glu Ala
275 280 285
Gln Val Lys Glu Ile Ala Arg Gly Leu Glu Gln Ser Gly His Arg Phe
290 295 300
Leu Trp Ser Leu Arg Arg Pro Pro Ser Glu Gln Thr Thr Thr Arg Ile
305 310 315 320
Pro Ser Asp Tyr Glu Asp Pro Ser Val Val Leu Pro Glu Gly Phe Leu
325 330 335
Asp Arg Thr Arg Gly Ile Gly Lys Val Ile Gly Trp Ala Pro Gln Val
340 345 350
Ala Val Leu Ala His Asp Ala Val Gly Gly Phe Val Ser His Cys Gly
355 360 365
Trp Asn Ser Leu Leu Glu Ser Leu Trp Phe Gly Val Pro Ser Ala Thr
370 375 380
Trp Pro Met Tyr Ala Glu Gln Gln Met Asn Ala Phe Glu Met Val Val
385 390 395 400
Asp Leu Gly Leu Ala Val Glu Ile Lys Leu Asp Tyr Glu Lys Asp Val
405 410 415
Phe Asn Pro Phe Asn Pro Lys Ala Asn Lys Ile Ile Asn Val Thr Ala
420 425 430
Gly Glu Ile Glu Ser Gly Met Arg Arg Val Met Glu Asp Asn Glu Val
435 440 445
Arg Val Arg Val Lys Glu Met Ser Ala Lys Ser Arg Ala Ala Val Val
450 455 460
Glu Gly Gly Ser Ser Tyr Ala Phe Val Gly Arg Leu Ile Gln Asp Phe
465 470 475 480
Ile Arg Asp
<210> 196
<211> 1452
<212> DNA
<213> 向日葵
<400> 196
atggcaaatg cagttgcaga actgattttt atcccgacac ctggtctggg tcatattatg 60
agcaccattg aactggcaaa actgctggtt aatcgtgatc agcgtctggc aattaccgtt 120
ctggttatta aaccgcctgg tatgaccagc ggtagcgcaa ttaccaccta tattgaaagc 180
ctgaccgaaa ccaccatgga tcgtattagc tttattcagc tgccgcaggt tgaaagcagc 240
ccgacacatg gtggtccgac cgaatttatt cgtagccata gcaaatatgt tcgtaatgcc 300
gttgttgatc tgcgtagcca gagcggtagc tgtcaggttg ttggttttgt tgttgatatg 360
ttttgcacca gcatgattga tgtggccaat gaatttaatg ttccgacctt tgtgtttttc 420
accagtagcg cagcatttct gggttttacc ctgtttatca aactgctgtg tgatgatctg 480
aatcgtgatg ttgttgaact gagcaatagc gataccgaaa tttcagtgcc gagctttgtt 540
aaaccggttc cgaccaaagt tttttggagc ctggttaaaa cccgtgaagg tctggatagc 600
gttcagcgcc tggcgaaaaa actgggtgaa gcaaaaggta ttatcgtgaa cacctttctg 660
gatctggaaa cccatgcaat tgaaagtctg agcgcagata ttagcattcc tccggtttat 720
ccggttggtc cgattctgaa cctggaaggt ggtagcggtg gtggtaaacc gtttgatgat 780
gatgttattc gttggctgga tagccagcct ccgagcagcg ttgtttttct gtgttttggt 840
agcatgggta gctttgatga agcacaggtt aaagaaattg cacgtggtct ggaacagagc 900
ggtcatcgtt ttctgtggtc actgcgtcgt ccgcctagcg aacagaccac cacacgtatt 960
ccgagcgatt atgaagatcc gagcgttgtt ctgccggaag gtttcctgga tcgtacccgt 1020
ggtattggta aagttattgg ttgggcacct caggttgcag ttctggcaca tgatgcagtt 1080
ggtggctttg ttagccattg tggttggaat agcctgctgg aaagcctgtg gtttggtgtt 1140
ccgagcgcaa cctggccgat gtatgcagaa cagcagatga atgcatttga aatggttgtg 1200
gatctgggtt tagccgtgga aattaaactg gattatgaga aggatgtgtt taacccgttt 1260
aatccgaaag ccaacaaaat cattaatgtg accgcaggcg aaattgaaag cggtatgcgt 1320
cgtgttatgg aagataatga agttcgtgtt cgcgtgaaag aaatgagcgc aaaaagccgt 1380
gcagcagttg ttgaaggtgg ttcaagctat gcatttgttg gtcgtctgat tcaggatttt 1440
atccgcgatt aa 1452
<210> 197
<211> 507
<212> PRT
<213> A. commosus
<400> 197
Met Lys Asp Val Thr Pro His Phe Val Leu Val Pro Leu Ala Ala Gln
1 5 10 15
Gly His Met Ile Pro Met Val Asp Met Ala Arg Leu Leu Ala Glu Arg
20 25 30
Gly Val Arg Val Thr Leu Ile Thr Thr Pro Val Asn Ala Ala Arg Ile
35 40 45
Arg Thr Ile Ile Asp Arg Val Arg Arg Ser Asn Leu Pro Val Glu Phe
50 55 60
Val Glu Leu Arg Phe Pro Cys Ala Glu Phe Gly Leu Pro Glu Gly Ser
65 70 75 80
Glu Asn Ile Asp Leu Leu Ser Thr Leu Glu His Tyr Lys Ala Phe Phe
85 90 95
Asp Ala Met Lys Leu Leu Lys Glu Pro Ile Glu Ala Leu Leu Arg Ser
100 105 110
Gln His Arg Arg Pro Asp Cys Met Ile Ala Asp Met Cys Asn Gly Trp
115 120 125
Thr Lys Asp Val Ala Arg Arg Leu Gly Ile Pro Arg Leu Leu Phe His
130 135 140
Gly Pro Ser Cys Phe Tyr Ile Leu Cys Ala Tyr Asn Met Ala Gln His
145 150 155 160
Arg Val Tyr Asp Arg Val Thr His Glu Phe Glu Pro Val Val Val Pro
165 170 175
Asp Val Pro Val Glu Val Val Thr Asn Lys Ala Glu Ser Pro Gly Phe
180 185 190
Phe Asn Trp Ser Gly Trp Glu Asp Leu Arg Ala Glu Val Leu Glu Ala
195 200 205
Glu Ser Thr Ala Asp Gly Val Val Ile Asn Thr Phe Tyr Asp Leu Glu
210 215 220
Pro Ser Phe Val Asp Cys Tyr Glu Lys Ile Met Gln Lys Lys Val Trp
225 230 235 240
Thr Val Gly Pro Leu Cys Leu Tyr Ser Lys Asp Val Asp Ser Lys Ala
245 250 255
Ala Arg Gly Asn Lys Ala Ala Val Asp His Arg Asp Ile Thr Thr Trp
260 265 270
Leu Asp Arg Lys Gly Ala Ser Ser Val Phe Tyr Val Ser Phe Gly Ser
275 280 285
Leu Val Leu Met Arg Pro Thr Gln Leu Ile Glu Ile Gly Lys Gly Leu
290 295 300
Leu Glu Cys Ser Asp His Arg Ser Phe Ile Trp Val Val Lys Glu Ala
305 310 315 320
Glu Leu Val Pro Glu Val Glu Lys Trp Leu Ser Glu Glu His Phe Ala
325 330 335
Glu Arg Thr Lys Glu Arg Gly Leu Leu Ile Lys Gly Trp Ala Pro Gln
340 345 350
Thr Val Ile Leu Leu His Pro Ala Ile Gly Gly Phe Leu Thr His Cys
355 360 365
Gly Trp Asn Ser Thr Leu Glu Ala Ile Ser Ala Gly Val Pro Met Leu
370 375 380
Thr Trp Pro His Phe Ala Asp Gln Phe Leu Asn Glu Lys Leu Val Val
385 390 395 400
Asp Val Leu Lys Ile Gly Arg Ser Leu Asp Val Lys Val Pro Arg Thr
405 410 415
His Val Thr Asp Asp Ser Thr Leu Leu Val Thr Lys Glu Lys Leu Arg
420 425 430
Lys Ala Val Ser Glu Leu Met Glu Gly Glu Glu Gly Glu Glu Met Arg
435 440 445
Arg Arg Ala Lys Ala Leu Ala Glu Lys Ala Lys Lys Ala Met Glu Glu
450 455 460
Gly Gly Ser Ser Tyr Arg Asn Met Asp Asp Met Ile Glu Cys Met Ala
465 470 475 480
Gly Arg Tyr Gly Glu Glu Glu Lys Val Glu Asp Ala Val Lys Glu Leu
485 490 495
Ser Asn Gly Phe Ser Ala His Val Val Val Thr
500 505
<210> 198
<211> 1524
<212> DNA
<213> A. commosus
<400> 198
atgaaagatg tgacaccgca ttttgttctg gttccgctgg cagcacaggg tcatatgatt 60
ccgatggttg atatggcacg tctgctggca gaacgtggtg ttcgtgttac cctgattacc 120
acaccggtta atgcagcacg tattcgtacc attattgatc gtgttcgtcg tagcaatctg 180
ccggttgaat ttgttgaact gcgttttccg tgtgcagaat ttggtctgcc ggaaggtagc 240
gaaaatattg atctgctgag caccctggaa cactataaag cattttttga tgccatgaaa 300
ctgctgaaag aaccgattga agcactgctg cgtagccagc atcgtcgtcc ggattgtatg 360
attgcagata tgtgtaatgg ttggaccaaa gatgttgcac gtcgtctggg tattccgcgt 420
ctgctgtttc atggtccgag ctgcttttat atcctgtgtg cctataatat ggcacagcat 480
cgtgtttatg atcgtgtgac ccatgaattt gaaccggttg ttgttccgga tgttccggtt 540
gaagtggtta ccaataaagc agaaagtccg ggttttttca attggagcgg ttgggaagat 600
ctgcgtgcag aagttctgga agccgaaagc accgcagatg gtgttgtgat taataccttt 660
tatgatctgg aaccgagctt cgttgattgc tatgaaaaaa tcatgcagaa aaaggtttgg 720
accgttggtc cgctgtgtct gtatagcaaa gatgtggata gcaaagcagc acgtggtaat 780
aaagccgcag ttgatcatcg tgacattacc acctggctgg atcgtaaagg tgcaagcagc 840
gttttttatg ttagctttgg tagcctggtt ctgatgcgtc cgacacagct gattgaaatt 900
ggtaaaggtc tgctggaatg cagcgatcat cgtagcttta tttgggttgt taaagaagca 960
gaactggttc cggaagttga aaaatggctg agcgaagaac attttgcaga acgtaccaaa 1020
gaacgcggtc tgctgattaa aggttgggct ccgcagaccg ttattctgct gcatccggca 1080
attggtggtt ttctgaccca ttgtggttgg aatagtaccc tggaagcaat tagtgccggt 1140
gttccgatgc tgacctggcc tcattttgcc gatcagtttc tgaatgaaaa actggttgtt 1200
gacgtgctga aaattggtcg tagcctggat gttaaagttc cgcgtacaca tgttaccgat 1260
gatagcaccc tgctggtgac caaagaaaaa ctgcgtaaag cagttagcga actgatggaa 1320
ggtgaagagg gtgaagaaat gcgtcgtcgt gcaaaagcac tggccgaaaa agcaaaaaaa 1380
gccatggaag aaggtggtag cagctatcgt aatatggatg atatgattga atgcatggca 1440
ggtcgttatg gcgaagaaga aaaagttgag gacgcagtta aagaactgag caatggtttt 1500
agcgcacatg ttgttgttac ctaa 1524
<210> 199
<211> 484
<212> PRT
<213> 番木瓜
<400> 199
Met Thr Gly Glu Leu Ile Phe Ile Pro Met Pro Ser Leu Ser His Ile
1 5 10 15
Ala Ser Thr Met Glu Ile Ala Lys Leu Leu Val His Arg Asp Asp Arg
20 25 30
Leu Ser Ile Thr Val Leu Leu Ile Ser Ser Gln Tyr Thr Thr Ser Ile
35 40 45
Thr Thr Tyr Ile Asn Ser Leu Ile Ala Ser Ser Asp Tyr Asp Arg Ile
50 55 60
Arg Phe Ile His Leu Pro Glu Leu Asp Ser Glu Glu Glu Pro Lys Arg
65 70 75 80
Pro Phe Met Ser Val Ile Asp Asp Asn Lys Pro Ile Val Lys Glu Ala
85 90 95
Val Thr Asn Leu Ala Leu Ser Phe Asp Pro Ser His Arg Leu Ala Gly
100 105 110
Phe Val Ile Asp Met Phe Cys Val Gly Met Ile Glu Val Ala Asp Glu
115 120 125
Leu Gly Leu Pro Ser Tyr Pro Phe Phe Thr Ser Ser Thr Ser Phe Leu
130 135 140
Ala Leu Gln Phe His Val Gln Thr Leu Ala Asp Glu Glu Glu Val Asp
145 150 155 160
Ile Thr Glu Phe Lys Asn Ser Asp Val Met Leu Pro Ile Pro Gly Leu
165 170 175
Val Asn Pro Leu Pro Ala Lys Thr Ile Leu Pro Ser Ala Met Leu Asn
180 185 190
Lys Asp Trp Leu Pro Tyr Val Leu Asn Gly Ala Arg Gly Phe Arg Lys
195 200 205
Thr Lys Gly Ile Met Val Asn Ser Phe Ala Glu Ile Glu Ser Asn Ala
210 215 220
Val Thr Ser Leu Ser Asn Ser Thr Val Pro Pro Val Tyr Thr Val Gly
225 230 235 240
Pro Ile Ile Asn Phe Lys Gly Asp Gly Gln Asp Ser Asp Thr Cys Thr
245 250 255
Ala His Lys Tyr Ser Asn Ile Met Thr Trp Leu Asp Asp Gln Pro Pro
260 265 270
Ser Ser Val Leu Phe Leu Cys Phe Gly Ser Leu Gly Ser Phe Asp Glu
275 280 285
Glu Gln Val Lys Glu Ile Ala Arg Ala Leu Glu Gly Ser Gly His Arg
290 295 300
Phe Leu Trp Ser Leu Arg Arg Pro Pro Pro Lys Asp Lys Thr Met Ser
305 310 315 320
Phe Pro Thr Glu Tyr Glu Asn Phe Glu Glu Val Leu Pro Glu Gly Phe
325 330 335
Val Asp Arg Thr Val Gly Met Gly Lys Val Met Gly Trp Ala Pro Gln
340 345 350
Val Ala Val Leu Ala His Pro Ser Ile Gly Gly Phe Val Thr His Cys
355 360 365
Gly Trp Asn Ser Ile Leu Glu Ser Val Trp Phe Gly Val Pro Met Ala
370 375 380
Ala Trp Pro Leu Tyr Ala Glu Gln Gln Phe Asn Ala Phe His Met Val
385 390 395 400
Val Glu Leu Gly Leu Ala Val Glu Ile Lys Met Asp Tyr Arg Lys Asp
405 410 415
Tyr Ala Ile Leu Gly Leu Gln Glu Glu Arg Val Ser Ala Glu Val Ile
420 425 430
Glu Lys Gly Ile Arg Cys Leu Met Glu Glu Asp Asn Asp Ala Arg Lys
435 440 445
Lys Val Lys Glu Met Ser Glu Ile Ser Arg Lys Ala Leu Met Asp Gly
450 455 460
Gly Ser Ser His Ala Val Leu Gly Gln Phe Ile Glu Asp Val Met Asn
465 470 475 480
Asn Ile Ser Ala
<210> 200
<211> 1455
<212> DNA
<213> 番木瓜
<400> 200
atgaccggtg aactgatttt tatcccgatg ccgagcctga gccatattgc aagcaccatg 60
gaaattgcaa aactgctggt tcatcgtgat gatcgtctga gcattaccgt tctgctgatt 120
agcagccagt ataccacctc aattaccacc tatattaaca gcctgattgc cagcagcgat 180
tatgatcgta ttcgttttat tcatctgccg gaactggata gcgaagaaga accgaaacgt 240
ccgtttatga gcgtgattga tgataacaaa ccgatcgtta aagaagccgt taccaatctg 300
gcactgagct ttgatccgag ccatcgtctg gcaggttttg ttattgatat gttttgcgtg 360
ggcatgattg aagttgcaga tgaactgggt ctgccgagct atccgttttt taccagcagc 420
accagctttc tggccctgca gtttcatgtt cagaccctgg ccgatgaaga agaagttgat 480
attaccgagt ttaagaactc cgatgttatg ctgccgattc ctggtctggt taatccgctg 540
cctgcaaaaa ccattctgcc gagtgcaatg ctgaataaag attggctgcc gtatgttctg 600
aatggtgcac gtggttttcg taaaacgaaa ggcattatgg ttaacagctt tgccgaaatt 660
gaaagcaatg cagttaccag cctgagcaat agcaccgttc cgcctgttta taccgttggt 720
ccgattatta actttaaagg tgatggtcag gatagcgata cctgtaccgc acacaaatat 780
agcaatatta tgacctggct ggatgatcag cctccgagca gcgttctgtt tctgtgtttt 840
ggtagcctgg gtagctttga tgaagaacag gttaaagaaa ttgcacgtgc cctggaaggt 900
agcggtcatc gttttctgtg gtcactgcgt cgtccgcctc cgaaagataa aaccatgagc 960
tttccgaccg aatatgaaaa ctttgaagaa gtgctgccgg aaggttttgt ggatcgcacc 1020
gttggtatgg gtaaagttat gggttgggca ccgcaggttg cagttctggc acatccgagc 1080
attggtggtt ttgtgaccca ttgtggttgg aatagcattc tggaaagcgt ttggtttggt 1140
gttccgatgg cagcatggcc tctgtatgca gaacagcagt ttaatgcatt tcatatggtg 1200
gtggaactgg gtttagcagt ggaaatcaaa atggattatc gcaaagatta tgccattctg 1260
ggcctgcaag aagaacgcgt tagcgcagaa gttattgaaa aaggtattcg ttgtctgatg 1320
gaagaggata atgatgcccg taaaaaagtg aaagaaatga gcgaaattag ccgcaaagca 1380
ctgatggatg gtggtagcag ccatgccgtt ctgggtcagt ttattgaaga tgtgatgaat 1440
aacatcagcg cctaa 1455
<210> 201
<211> 470
<212> PRT
<213> 向日葵
<400> 201
Met Glu Arg Thr Pro His Ile Ala Ile Val Pro Ser Pro Gly Met Gly
1 5 10 15
His Leu Ile Pro Leu Val Glu Phe Ala Lys Arg Leu Lys Asn Asn His
20 25 30
Asn Ile Ser Ser Thr Phe Ile Ile Pro Asn Asp Gly Pro Leu Ser Ile
35 40 45
Ser Gln Lys Ala Phe Leu Asp Ser Leu Pro Met Gly Leu Asn His Ile
50 55 60
Ile Leu Pro Pro Val Asn Phe Asp Asp Leu Pro Gln Asp Thr Gln Met
65 70 75 80
Glu Thr Arg Ile Ser Leu Met Val Thr Arg Ser Leu Asp Ser Leu Arg
85 90 95
Glu Val Phe Lys Ser Leu Val Ala Glu His Asn Met Val Ala Leu Phe
100 105 110
Ile Asp Leu Phe Gly Thr Asp Ala Phe Asp Val Ala Ile Glu Phe Gly
115 120 125
Val Ser Pro Tyr Val Phe Phe Pro Ser Thr Ala Met Ala Leu Ser Leu
130 135 140
Phe Leu Tyr Leu Pro Lys Leu Asp Gln Met Thr Ser Cys Glu Tyr Arg
145 150 155 160
Asp Leu Pro Glu Pro Val Gln Ile Pro Gly Cys Leu Pro Val Arg Gly
165 170 175
Gln Asp Leu Leu Asp Pro Val Gln Asp Arg Lys Asn Asp Ala Tyr Lys
180 185 190
Trp Val Leu His Asn Ala Lys Arg Tyr Met Met Ala Glu Gly Ile Ala
195 200 205
Val Asn Ser Phe Lys Glu Leu Glu Gly Gly Ala Leu Lys Ala Leu Leu
210 215 220
Glu Ala Glu Pro Gly Lys Pro Lys Ile Tyr Pro Val Gly Pro Leu Ile
225 230 235 240
Gln Thr Gly Ser Ser Ser Asp Val Asp Gly Ser Gly Cys Leu Lys Trp
245 250 255
Leu Asp Gly Gln Pro Cys Gly Ser Val Leu Tyr Ile Ser Phe Gly Ser
260 265 270
Gly Gly Thr Leu Ser Ser Asn Gln Leu Asn Glu Leu Ala Met Gly Leu
275 280 285
Glu Leu Ser Glu Gln Arg Phe Ile Trp Val Val Arg Ser Pro Ser Asp
290 295 300
Gln Ala Asn Ala Thr Tyr Phe Asn Ser His Gly His Lys Asp Pro Leu
305 310 315 320
Gly Phe Leu Pro Lys Gly Phe Leu Glu Arg Thr Lys Gly Asn Gly Phe
325 330 335
Val Val Ser Ser Trp Ala Pro Gln Ala Gln Ile Leu Ser His Ser Ser
340 345 350
Thr Gly Gly Phe Leu Thr His Cys Gly Trp Asn Ser Ile Leu Glu Thr
355 360 365
Val Val His Gly Val Pro Val Ile Ala Trp Pro Leu Tyr Ala Glu Gln
370 375 380
Lys Met Asn Ala Val Ser Leu Thr Glu Gly Ile Lys Val Ala Leu Arg
385 390 395 400
Pro Thr Val Gly Glu Asn Gly Ile Ile Gly Arg Val Glu Ile Ala Arg
405 410 415
Val Val Lys Ser Leu Leu Glu Gly Glu Glu Gly Lys Ala Ile Arg Ser
420 425 430
Arg Ile Arg Asp Leu Lys Asp Ala Ala Ala Asn Val Ile Ser Lys Asp
435 440 445
Gly Cys Ser Thr Lys Thr Leu Asp Lys Leu Ala Ser Met Leu Lys Asn
450 455 460
Lys Asn Lys Leu Ser Leu
465 470
<210> 202
<211> 1413
<212> DNA
<213> 向日葵
<400> 202
atggaacgta caccgcatat tgcaattgtt ccgagtcctg gtatgggtca tctgattccg 60
ctggttgaat ttgcaaaacg cctgaaaaac aaccacaata ttagcagcac ctttatcatt 120
ccgaacgatg gtccgctgag cattagccag aaagcatttt tagatagcct gccgatgggt 180
ctgaaccata ttattctgcc tccggtgaat tttgatgatc tgccgcagga tacccagatg 240
gaaacccgta ttagcctgat ggttacccgt agcctggata gtctgcgtga agtgtttaaa 300
agcctggttg cagaacataa catggtggca ctgtttattg acctgtttgg caccgatgca 360
tttgatgttg caattgaatt tggtgttagc ccgtatgttt tttttccgag caccgcaatg 420
gcactgagcc tgtttctgta tctgccgaaa ctggatcaaa tgaccagctg tgaatatcgc 480
gatctgccgg aaccggtgca gattccgggt tgtctgccgg ttcgtggtca ggatctgctg 540
gatccggttc aggatcgtaa aaatgatgca tataaatggg tgctgcataa cgccaaacgt 600
tatatgatgg cagaaggtat tgccgtcaac agctttaaag aactggaagg tggtgcactg 660
aaagcactgc tggaagcaga accgggtaaa ccgaaaatct atccggttgg tcctctgatt 720
cagaccggta gcagcagtga tgttgatggt agcggttgtc tgaaatggct ggatggtcag 780
ccgtgtggta gcgttctgta tattagcttt ggtagtggtg gcaccctgag cagcaatcag 840
ctgaatgaac tggcaatggg tttagaactg agcgaacagc gttttatttg ggttgttcgt 900
agcccgagcg atcaggcaaa tgcaacctat tttaacagcc atggtcataa agatccgctg 960
ggttttctgc ctaaaggttt tctggaacgc accaaaggta atggttttgt tgttagcagc 1020
tgggcaccgc aggcacagat tctgagccat agcagtaccg gtggttttct gacccattgt 1080
ggctggaata gcattctgga aaccgttgtt catggtgttc cggttattgc atggcctctg 1140
tatgcagaac agaaaatgaa tgcagttagc ctgaccgaag gtattaaagt tgcactgcgt 1200
ccgaccgttg gtgaaaatgg tattattggt cgtgttgaaa ttgcccgtgt tgtgaaaagc 1260
ctgttagaag gtgaagaagg taaagcaatt cgtagccgta ttcgtgatct gaaagatgca 1320
gcagcaaatg tgattagcaa agatggttgt agcaccaaaa cactggataa actggcaagc 1380
atgctgaaga acaaaaacaa actgtccctg taa 1413
<210> 203
<211> 485
<212> PRT
<213> 潘那利番茄
<400> 203
Met Asp Lys Arg Ala Asp Gln Leu His Val Tyr Phe Leu Pro Met Met
1 5 10 15
Ala Pro Gly His Met Ile Pro Leu Val Asp Met Ala Arg Gln Phe Ser
20 25 30
Arg His Gly Val Lys Val Thr Ile Val Thr Thr Pro Leu Asn Ala Thr
35 40 45
Lys Phe Ser Lys Thr Ile Gln Lys Asp Arg Glu Phe Gly Ser Asp Ile
50 55 60
Cys Ile Arg Thr Thr Glu Phe Pro Cys Lys Glu Ala Gly Leu Pro Glu
65 70 75 80
Gly Cys Glu Asn Leu Ala Ser Thr Thr Thr Ser Glu Met Thr Met Lys
85 90 95
Phe Ile Lys Ala Leu Tyr Leu Phe Glu Gln Pro Val Glu Lys Phe Met
100 105 110
Glu Glu Asp His Pro Asp Cys Leu Val Ala Gly Thr Phe Phe Ala Trp
115 120 125
Ala Val Asp Val Ala Ala Lys Leu Gly Ile Pro Arg Leu Ala Phe Asn
130 135 140
Gly Thr Gly Leu Leu Pro Met Cys Ala Tyr Asn Cys Leu Met Glu His
145 150 155 160
Lys Pro His Leu Lys Val Glu Ser Glu Thr Glu Glu Phe Val Ile Pro
165 170 175
Gly Leu Pro Asp Thr Ile Lys Met Ser Arg Ser Lys Leu Ser Gln His
180 185 190
Trp Val Asp Glu Lys Glu Thr Pro Met Thr Pro Ile Ile Lys Asp Phe
195 200 205
Met Arg Ala Glu Ala Thr Ser Tyr Gly Ala Ile Val Asn Ser Phe Tyr
210 215 220
Glu Leu Glu Pro Asn Tyr Val Gln His Phe Arg Glu Val Val Gly Arg
225 230 235 240
Lys Val Trp His Val Gly Pro Val Ser Leu Cys Asn Lys Asp Asn Glu
245 250 255
Asp Lys Ser Gln Arg Gly Gln Asp Ser Ser Leu Ser Glu Gln Lys Cys
260 265 270
Leu Asp Trp Leu Asn Thr Lys Glu Pro Lys Ser Val Ile Tyr Ile Cys
275 280 285
Phe Gly Ser Met Ser Ile Phe Ser Ser Asp Gln Leu Leu Glu Ile Ala
290 295 300
Thr Ala Leu Glu Ala Ser Asp Gln Gln Phe Ile Trp Val Val Arg Gln
305 310 315 320
Asn Thr Thr Asn Glu Glu Gln Glu Lys Trp Met Pro Glu Gly Phe Glu
325 330 335
Glu Lys Val Asn Gly Arg Gly Leu Ile Ile Lys Gly Trp Ala Pro Gln
340 345 350
Val Leu Ile Leu Asp His Glu Ala Thr Gly Gly Phe Val Thr His Cys
355 360 365
Gly Trp Asn Ser Leu Leu Glu Gly Val Ser Ala Gly Val Pro Met Val
370 375 380
Thr Trp Pro Leu Ser Ala Glu Gln Phe Phe Asn Glu Lys Leu Leu Val
385 390 395 400
Glu Ile Leu Lys Ile Gly Val Pro Val Gly Val Gln Ala Trp Ser Gln
405 410 415
Arg Thr Asp Ser Arg Val Pro Ile Asn Arg Glu Asn Ile Leu Arg Ala
420 425 430
Val Thr Lys Leu Met Val Gly Gln Glu Ala Glu Glu Met Gln Gly Arg
435 440 445
Ala Ala Ala Leu Gly Lys Ser Ala Lys Met Ala Val Glu Lys Gly Gly
450 455 460
Ser Ser Asp Asn Ser Leu Val Ser Leu Leu Glu Glu Leu Arg Asn Gly
465 470 475 480
Lys Ser Ser Ser Asn
485
<210> 204
<211> 1458
<212> DNA
<213> 潘那利番茄
<400> 204
atggataaac gtgcagatca gctgcatgtt tattttctgc cgatgatggc accgggtcat 60
atgattccgc tggttgatat ggcacgtcag tttagccgtc atggtgttaa agttaccatt 120
gttaccacac cgctgaatgc aaccaaattt agcaaaacca ttcagaaaga tcgcgaattt 180
ggtagcgata tttgtattcg taccaccgaa tttccgtgta aagaagcagg tctgccggaa 240
ggttgtgaaa atctggcaag caccaccacc agtgaaatga ccatgaaatt tatcaaagcc 300
ctgtacctgt ttgaacagcc ggttgaaaaa ttcatggaag aagatcatcc ggattgtctg 360
gttgcaggca ccttttttgc atgggcagtt gatgttgcag caaaactggg tattccgcgt 420
ctggcattta atggtacagg tctgctgccg atgtgtgcat ataattgtct gatggaacat 480
aaaccgcacc tgaaagttga aagcgaaacc gaagaatttg ttattccggg tctgcctgat 540
acgattaaaa tgagccgtag caaactgagc cagcattggg ttgatgaaaa agaaaccccg 600
atgacaccga tcatcaaaga ttttatgcgt gccgaagcaa ccagctatgg tgcaattgtt 660
aatagctttt atgagctgga accgaactat gtgcagcatt ttcgtgaagt tgttggtcgt 720
aaagtttggc atgttggtcc ggttagcctg tgcaataaag ataatgaaga taaaagccag 780
cgtggtcagg atagcagcct gagcgaacag aaatgtctgg attggctgaa taccaaagaa 840
ccgaaaagcg tgatctatat ttgctttggt agcatgagca tctttagcag cgatcaactg 900
ctggaaattg caaccgcact ggaagcaagc gatcagcagt ttatttgggt tgttcgtcag 960
aataccacca acgaagaaca agaaaaatgg atgcctgaag gctttgaaga aaaagttaat 1020
ggtcgtggcc tgattatcaa aggttgggca ccgcaggttc tgattctgga tcatgaagca 1080
accggtggtt ttgttaccca ttgtggttgg aatagcctgc tggaaggtgt tagtgccggt 1140
gttccgatgg ttacctggcc tctgagcgca gaacagtttt ttaacgaaaa actgctggtc 1200
gagattctga aaattggtgt tccggttggt gttcaggcat ggtcacagcg taccgatagc 1260
cgtgttccta ttaatcgtga aaatattctg cgtgccgtta ccaaactgat ggttggtcaa 1320
gaggccgaag aaatgcaggg tcgtgcagca gcactgggta aaagcgcaaa aatggcagtt 1380
gaaaaaggtg gcagcagcga taatagcctg gttagcttac tggaagaact gcgtaatggt 1440
aaaagcagca gcaactaa 1458
<210> 205
<211> 471
<212> PRT
<213> S. pennellii
<400> 205
Met Ala Gln Ile Pro His Ile Ala Ile Leu Pro Ser Pro Gly Met Gly
1 5 10 15
His Leu Ile Pro Leu Val Glu Phe Ala Lys Arg Ile Phe Leu His His
20 25 30
Gln Phe Ser Val Ser Leu Ile Leu Pro Thr Asp Gly Pro Ile Ser Asn
35 40 45
Ala Gln Lys Ile Phe Leu Asn Ser Leu Pro Ser Ser Met Asp Tyr His
50 55 60
Leu Leu Pro Pro Val Asn Phe Asp Asp Leu Pro Glu Asp Val Lys Ile
65 70 75 80
Glu Thr Arg Ile Ser Leu Thr Val Ser Arg Ser Leu Thr Ser Leu Arg
85 90 95
Gln Val Leu Asp Ser Ile Ile Glu Ser Lys Arg Thr Val Ala Leu Val
100 105 110
Val Asp Leu Phe Gly Thr Asp Ala Phe Asp Val Ala Ile Asp Leu Lys
115 120 125
Ile Ser Pro Tyr Ile Phe Phe Pro Ser Thr Ala Met Ala Leu Ser Leu
130 135 140
Phe Leu Tyr Leu Pro Asn Leu Asp Glu Thr Val Ser Cys Glu Tyr Arg
145 150 155 160
Asp Leu Pro Asp Pro Ile Gln Ile Pro Gly Cys Thr Pro Ile His Gly
165 170 175
Lys Asp Leu Leu Asp Pro Val Gln Asp Arg Asn Asp Glu Ser Tyr Lys
180 185 190
Trp Leu Leu His His Val Lys Arg Tyr Gly Met Ala Glu Gly Ile Ile
195 200 205
Val Asn Ser Phe Lys Glu Leu Glu Gly Gly Ala Ile Gly Ala Leu Gln
210 215 220
Lys Asp Glu Pro Gly Lys Pro Thr Val Tyr Pro Val Gly Pro Leu Ile
225 230 235 240
Gln Met Asp Ser Gly Ser Lys Val Asp Gly Ser Glu Cys Met Thr Trp
245 250 255
Leu Asp Glu Gln Pro Arg Gly Ser Val Leu Tyr Ile Ser Tyr Gly Ser
260 265 270
Gly Gly Thr Leu Ser His Glu Gln Leu Ile Glu Val Ala Ala Gly Leu
275 280 285
Glu Met Ser Glu Gln Arg Phe Leu Trp Val Val Arg Cys Pro Asn Asp
290 295 300
Lys Ile Ala Asn Ala Thr Phe Phe Asn Val Gln Asp Ser Thr Asn Pro
305 310 315 320
Leu Glu Phe Leu Pro Lys Gly Phe Leu Glu Arg Thr Lys Gly Phe Gly
325 330 335
Leu Val Leu Pro Asn Trp Ala Pro Gln Ala Arg Ile Leu Ser His Glu
340 345 350
Ser Thr Gly Gly Phe Leu Thr His Cys Gly Trp Asn Ser Thr Leu Glu
355 360 365
Ser Val Val His Gly Val Pro Leu Ile Ala Trp Pro Leu Tyr Ala Glu
370 375 380
Gln Lys Met Asn Ala Val Met Leu Ser Glu Asp Ile Lys Val Ala Leu
385 390 395 400
Arg Pro Lys Val Asn Glu Glu Asn Gly Ile Val Gly Arg Leu Glu Ile
405 410 415
Ala Lys Val Val Lys Gly Leu Met Glu Gly Glu Glu Gly Lys Gly Val
420 425 430
Arg Ser Arg Met Arg Asp Leu Lys Asp Ala Ala Ala Lys Val Leu Ser
435 440 445
Glu Asp Gly Ser Ser Thr Lys Ala Leu Ala Glu Leu Ala Thr Lys Leu
450 455 460
Lys Lys Lys Val Ser Asn Asn
465 470
<210> 206
<211> 1416
<212> DNA
<213> 潘那利番茄
<400> 206
atggcacaga ttccgcatat tgcaattctg ccgagtcctg gtatgggtca tctgattccg 60
ctggttgaat ttgccaaacg tatttttctg catcaccagt ttagcgttag cctgatcctg 120
ccgaccgatg gtccgattag caatgcacag aaaatctttc tgaatagcct gccgagcagc 180
atggattatc atctgctgcc tccggttaat tttgatgatc tgccggaaga tgtgaaaatt 240
gaaacccgta ttagcctgac cgttagccgt agtctgacca gcctgcgtca ggttctggat 300
agcattattg aaagcaaacg taccgttgca ctggttgttg acctgtttgg caccgatgca 360
tttgatgttg caattgatct gaaaatcagc ccgtatatct tttttccgag caccgcaatg 420
gcactgagcc tgtttctgta tctgccgaat ctggatgaaa ccgttagctg tgaatatcgt 480
gatctgcctg atccgattca gattccgggt tgtaccccga ttcatggtaa agatctgctg 540
gatccggtgc aggatcgtaa tgatgaaagc tataaatggc tgctgcatca cgttaaacgt 600
tatggtatgg cagaaggcat tatcgtcaac agctttaaag aactggaagg tggtgcaatt 660
ggtgcactgc agaaagatga accgggtaaa ccgaccgttt atccggttgg tccgctgatt 720
cagatggata gcggtagcaa agttgatggt agcgaatgta tgacctggct ggatgaacag 780
cctcgtggta gcgttctgta tattagctat ggtagcggtg gcaccctgag ccatgaacag 840
ctgattgaag ttgcagcagg tctggaaatg agcgaacagc gttttctgtg ggttgttcgt 900
tgtccgaatg ataaaattgc aaacgccacc ttttttaacg ttcaggatag caccaatccg 960
ctggaatttc tgccgaaagg ttttctggaa cgtaccaaag gttttggtct ggtgctgccg 1020
aattgggcac cgcaggcacg tattctgagt catgaaagca ccggtggttt tctgacccat 1080
tgtggttgga atagcaccct ggaaagcgtt gttcatggtg tgccgctgat tgcatggcct 1140
ctgtatgcag aacagaaaat gaatgcagtt atgctgagcg aggatattaa agttgcactg 1200
cgtccgaaag tgaatgaaga aaatggtatt gttggtcgcc tggaaattgc caaagttgtt 1260
aaaggtctga tggaaggtga agaaggtaaa ggcgttcgta gccgtatgcg cgatctgaaa 1320
gatgccgcag caaaagttct gagcgaagat ggtagcagca ccaaagcact ggcagaactg 1380
gcaaccaaac tgaaaaaaaa ggtcagcaac aattaa 1416
<210> 207
<211> 480
<212> PRT
<213> 番红花
<400> 207
Met Gly Ser Glu Gly Arg Gln Leu His Ile Phe Met Phe Pro Phe Met
1 5 10 15
Ala His Gly His Met Ile Pro Ile Val Asp Met Ala Lys Leu Phe Ala
20 25 30
Ser Arg Gly Ile Lys Ile Thr Ile Val Thr Thr Pro Leu Asn Ser Ile
35 40 45
Ser Ile Ser Lys Ser Leu His Asn Cys Ser Pro Asn Ser Leu Ile Gln
50 55 60
Leu Leu Ile Leu Lys Phe Pro Ala Ala Glu Ala Gly Leu Pro Asp Gly
65 70 75 80
Cys Glu Asn Ala Asp Ser Ile Pro Ser Met Asp Leu Leu Pro Lys Phe
85 90 95
Phe Glu Ala Val Ser Leu Leu Gln Pro Pro Phe Glu Glu Ala Leu His
100 105 110
Asn Asn Arg Pro Asp Cys Leu Ile Ser Asp Met Phe Phe Pro Trp Thr
115 120 125
Asn Asp Val Ala Asp Arg Val Gly Ile Pro Arg Leu Ile Phe His Gly
130 135 140
Thr Ser Cys Phe Ser Leu Cys Ser Ser Glu Phe Met Arg Leu His Lys
145 150 155 160
Pro Tyr Gln His Val Ser Ser Asp Thr Glu Pro Phe Thr Ile Pro Tyr
165 170 175
Leu Pro Gly Asp Ile Lys Leu Thr Lys Met Lys Leu Pro Ile Phe Val
180 185 190
Arg Glu Asn Ser Glu Asn Glu Phe Ser Lys Phe Ile Thr Lys Val Lys
195 200 205
Glu Ser Glu Ser Phe Cys Tyr Gly Val Val Val Asn Ser Phe Tyr Glu
210 215 220
Leu Glu Ala Glu Tyr Val Asp Cys Tyr Lys Asp Val Leu Gly Arg Lys
225 230 235 240
Thr Trp Thr Ile Gly Pro Leu Ser Leu Thr Asn Thr Lys Thr Gln Glu
245 250 255
Ile Thr Leu Arg Gly Arg Glu Ser Ala Ile Asp Glu His Glu Cys Leu
260 265 270
Lys Trp Leu Asp Ser Gln Lys Pro Asn Ser Val Val Tyr Val Cys Phe
275 280 285
Gly Ser Leu Ala Lys Phe Asn Ser Ala Gln Leu Lys Glu Ile Ala Ile
290 295 300
Gly Leu Glu Ala Ser Gly Lys Lys Phe Ile Trp Val Val Arg Lys Gly
305 310 315 320
Lys Gly Glu Glu Glu Glu Glu Glu Gln Asn Trp Leu Pro Glu Gly Tyr
325 330 335
Glu Glu Arg Met Glu Gly Thr Gly Leu Ile Ile Arg Gly Trp Ala Pro
340 345 350
Gln Val Leu Ile Leu Asp His Pro Ser Val Gly Gly Phe Val Thr His
355 360 365
Cys Gly Trp Asn Ser Thr Leu Glu Gly Val Ala Ala Gly Val Pro Met
370 375 380
Val Thr Trp Pro Val Gly Ala Glu Gln Phe Tyr Asn Glu Lys Leu Val
385 390 395 400
Thr Glu Val Leu Lys Thr Gly Val Gly Val Gly Val Gln Lys Trp Ala
405 410 415
Pro Gly Val Gly Asp Phe Ile Glu Ser Glu Ala Val Glu Lys Ala Ile
420 425 430
Arg Arg Ile Met Glu Lys Glu Gly Glu Glu Met Arg Asn Arg Ala Ile
435 440 445
Glu Leu Gly Lys Lys Ala Lys Trp Ala Val Gly Glu Glu Gly Ser Ser
450 455 460
Tyr Ser Asn Leu Asp Ala Leu Ile Glu Glu Leu Lys Ser Leu Ala Phe
465 470 475 480
<210> 208
<211> 1443
<212> DNA
<213> 番红花
<400> 208
atgggttctg aaggtagaca attgcacatt ttcatgttcc cattcatggc tcatggtcat 60
atgattccaa tagttgatat ggctaagttg ttcgcctcaa gaggtattaa gattaccatc 120
gttactacgc ccttgaactc catttctatc tctaagtcat tgcacaactg ctccccaaat 180
tctttgattc agttgctgat tttgaagttc ccagctgctg aagctggttt gccagatggt 240
tgtgaaaatg ctgattctat cccatctatg gacttgttgc caaagttttt cgaagccgtt 300
tctttgttgc aaccaccatt tgaagaagcc ttgcataaca atagaccaga ctgcttgatt 360
tccgatatgt tttttccatg gaccaacgat gttgctgata gagttggtat tccaagattg 420
atcttccatg gcacctcttg cttttctttg tgttcttctg aattcatgag gctgcataag 480
ccataccaac atgtttcttc agatactgag ccattcacca ttccatattt gccaggtgat 540
attaagctga ccaaaatgaa gttgccaatc ttcgtcagag aaaactccga aaacgaattc 600
tccaagttca tcaccaaggt caaagaatct gaatctttct gctacggtgt tgtcgttaac 660
tctttctatg aattggaagc cgaatacgtt gattgctaca aagatgtttt gggtagaaag 720
acttggacta tcggtccatt gtctttgact aacactaaga cccaagaaat caccttgaga 780
ggtagagaat ctgccattga tgaacatgaa tgtttgaagt ggttggactc tcaaaagcca 840
aactctgttg tttacgtttg ctttggttct ttggccaagt ttaactccgc tcagttgaaa 900
gaaattgcta ttggtttgga agcctccggt aagaagttta tttgggttgt tagaaaaggt 960
aagggcgaag aagaagagga agaacaaaat tggttgccag aaggttacga agaaagaatg 1020
gaaggtactg gtttgattat tagaggttgg gctccacaag ttttgatttt ggatcatcca 1080
tctgttggtg gtttcgttac tcattgtggt tggaattcta ctttggaagg tgttgctgct 1140
ggtgttccaa tggttacttg gccagttggt gctgaacaat tttacaacga aaagttggtt 1200
accgaggtct tgaaaactgg tgttggtgta ggtgttcaaa aatgggctcc aggtgtcggt 1260
gattttattg aatctgaagc tgttgagaag gccatcagac gtattatgga aaaagaaggt 1320
gaagagatga gaaacagagc cattgaattg ggtaaaaaag ctaaatgggc tgtcggtgaa 1380
gaaggttctt cttactctaa tttggatgcc ttgatcgaag agttgaagtc tttggctttc 1440
taa 1443
<210> 209
<211> 805
<212> PRT
<213> 大豆
<400> 209
Met Ala Thr Asp Arg Leu Thr Arg Val His Ser Leu Arg Glu Arg Leu
1 5 10 15
Asp Glu Thr Leu Thr Ala Asn Arg Asn Glu Ile Leu Ala Leu Leu Ser
20 25 30
Arg Ile Glu Ala Lys Gly Lys Gly Ile Leu Gln His His Gln Val Ile
35 40 45
Ala Glu Phe Glu Glu Ile Pro Glu Glu Asn Arg Gln Lys Leu Thr Asp
50 55 60
Gly Ala Phe Gly Glu Val Leu Arg Ser Thr Gln Glu Ala Ile Val Leu
65 70 75 80
Pro Pro Trp Val Ala Leu Ala Val Arg Pro Arg Pro Gly Val Trp Glu
85 90 95
Tyr Leu Arg Val Asn Val His Ala Leu Val Val Glu Glu Leu Gln Pro
100 105 110
Ala Glu Tyr Leu His Phe Lys Glu Glu Leu Val Asp Gly Ser Ser Asn
115 120 125
Gly Asn Phe Val Leu Glu Leu Asp Phe Glu Pro Phe Asn Ala Ala Phe
130 135 140
Pro Arg Pro Thr Leu Asn Lys Ser Ile Gly Asn Gly Val Gln Phe Leu
145 150 155 160
Asn Arg His Leu Ser Ala Lys Leu Phe His Asp Lys Glu Ser Leu His
165 170 175
Pro Leu Leu Glu Phe Leu Arg Leu His Ser Val Lys Gly Lys Thr Leu
180 185 190
Met Leu Asn Asp Arg Ile Gln Asn Pro Asp Ala Leu Gln His Val Leu
195 200 205
Arg Lys Ala Glu Glu Tyr Leu Gly Thr Val Pro Pro Glu Thr Pro Tyr
210 215 220
Ser Glu Phe Glu His Lys Phe Gln Glu Ile Gly Leu Glu Arg Gly Trp
225 230 235 240
Gly Asp Asn Ala Glu Arg Val Leu Glu Ser Ile Gln Leu Leu Leu Asp
245 250 255
Leu Leu Glu Ala Pro Asp Pro Cys Thr Leu Glu Thr Phe Leu Gly Arg
260 265 270
Ile Pro Met Val Phe Asn Val Val Ile Leu Ser Pro His Gly Tyr Phe
275 280 285
Ala Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val
290 295 300
Tyr Ile Leu Asp Gln Val Arg Ala Leu Glu Asn Glu Met Leu His Arg
305 310 315 320
Ile Lys Gln Gln Gly Leu Asp Ile Val Pro Arg Ile Leu Ile Ile Thr
325 330 335
Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg Leu Glu
340 345 350
Lys Val Phe Gly Thr Glu His Ser His Ile Leu Arg Val Pro Phe Arg
355 360 365
Thr Glu Lys Gly Ile Val Arg Lys Trp Ile Ser Arg Phe Glu Val Trp
370 375 380
Pro Tyr Leu Glu Thr Tyr Thr Glu Asp Val Ala His Glu Leu Ala Lys
385 390 395 400
Glu Leu Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser Asp Gly
405 410 415
Asn Ile Val Ala Ser Leu Leu Ala His Lys Leu Gly Val Thr Gln Cys
420 425 430
Thr Ile Ala His Ala Leu Glu Lys Thr Lys Tyr Pro Glu Ser Asp Ile
435 440 445
Tyr Trp Lys Lys Leu Glu Glu Arg Tyr His Phe Ser Cys Gln Phe Thr
450 455 460
Ala Asp Leu Phe Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr
465 470 475 480
Phe Gln Glu Ile Ala Gly Ser Lys Asp Thr Val Gly Gln Tyr Glu Ser
485 490 495
His Thr Ala Phe Thr Leu Pro Gly Leu Tyr Arg Val Val His Gly Ile
500 505 510
Asp Val Phe Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Gln
515 520 525
Thr Ile Tyr Phe Pro His Thr Glu Thr Ser Arg Arg Leu Thr Ser Phe
530 535 540
His Pro Glu Ile Glu Glu Leu Leu Tyr Ser Ser Val Glu Asn Glu Glu
545 550 555 560
His Ile Cys Val Leu Lys Asp Arg Ser Lys Pro Ile Ile Phe Thr Met
565 570 575
Ala Arg Leu Asp Arg Val Lys Asn Ile Thr Gly Leu Val Glu Trp Tyr
580 585 590
Gly Lys Asn Ala Lys Leu Arg Glu Leu Val Asn Leu Val Val Val Ala
595 600 605
Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met
610 615 620
Lys Lys Met Tyr Gly Leu Ile Glu Thr Tyr Lys Leu Asn Gly Gln Phe
625 630 635 640
Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr
645 650 655
Arg Val Ile Cys Asp Thr Arg Gly Ala Phe Val Gln Pro Ala Val Tyr
660 665 670
Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro
675 680 685
Thr Phe Ala Thr Cys Asn Gly Gly Pro Ala Glu Ile Ile Val His Gly
690 695 700
Lys Ser Gly Phe His Ile Asp Pro Tyr His Gly Asp Arg Ala Ala Asp
705 710 715 720
Leu Leu Val Asp Phe Phe Glu Lys Cys Lys Leu Asp Pro Thr His Trp
725 730 735
Asp Lys Ile Ser Lys Ala Gly Leu Gln Arg Ile Glu Glu Lys Tyr Thr
740 745 750
Trp Gln Ile Tyr Ser Gln Arg Leu Leu Thr Leu Thr Gly Val Tyr Gly
755 760 765
Phe Trp Lys His Val Ser Asn Leu Asp Arg Arg Glu Ser Arg Arg Tyr
770 775 780
Leu Glu Met Phe Tyr Ala Leu Lys Tyr Arg Lys Leu Ala Glu Ser Val
785 790 795 800
Pro Leu Ala Ala Glu
805
<210> 210
<211> 2418
<212> DNA
<213> 大豆
<400> 210
atggcaaccg atcgtctgac ccgtgttcat agcctgcgtg aacgtctgga tgaaaccctg 60
accgcaaatc gtaatgaaat tctggcactg ctgagccgta ttgaagcaaa aggtaaaggt 120
attctgcagc atcatcaggt gattgccgaa tttgaagaaa ttccggaaga aaatcgtcag 180
aaactgaccg atggtgcatt tggtgaagtt ctgcgtagca cccaagaagc aattgttctg 240
cctccgtggg ttgcactggc agttcgtccg cgtcctggtg tttgggaata tctgcgtgtt 300
aatgttcatg cactggttgt tgaagaactg cagcctgcag agtatctgca ttttaaagaa 360
gaactggtag acggtagcag caatggtaat tttgttctgg aactggattt tgagccgttt 420
aatgcagcat ttccgcgtcc gacactgaat aaaagcattg gtaatggtgt tcagttcctg 480
aatcgtcatc tgagcgcaaa actgtttcat gataaagaaa gcctgcatcc gctgctggaa 540
tttctgcgtc tgcatagcgt taaaggtaaa accctgatgc tgaatgatcg tattcagaat 600
ccggatgcac tgcagcatgt gctgcgtaaa gcagaagaat atctgggcac cgttccgcct 660
gaaacaccgt atagtgaatt tgaacacaag tttcaagaaa tcggtctgga acgtggttgg 720
ggtgataatg cagaacgtgt gctggaaagc attcagctgc tgctggatct gctggaagca 780
ccggatccgt gtacactgga aacctttctg ggtcgtattc cgatggtttt taatgtggtt 840
attctgagtc cgcatggtta ttttgcacag gataatgttc tgggttatcc tgataccggt 900
ggtcaggttg tttatattct ggatcaggtt cgtgcactgg aaaatgagat gctgcatcgt 960
attaaacagc aaggcctgga tattgttccg cgtattctga ttattacccg tctgctgccg 1020
gatgcagttg gcaccacctg tggtcagcgt ctggaaaaag tttttggcac cgaacatagc 1080
catattctgc gtgtgccgtt tcgtaccgaa aaaggtattg ttcgtaaatg gattagccgc 1140
tttgaagttt ggccgtatct ggaaacatat accgaagatg ttgcacatga actggcaaaa 1200
gagctgcagg gtaaaccgga tctgattgtt ggtaattata gcgacggtaa tattgttgca 1260
agcctgctgg cacataaact gggtgttacc cagtgtacca ttgcacatgc cctggaaaaa 1320
accaaatatc cggaaagcga tatctactgg aagaagctgg aagaacgtta tcattttagc 1380
tgtcagttta ccgcagacct gtttgcaatg aatcataccg attttatcat caccagcacc 1440
tttcaagaga ttgcaggtag caaagatacc gtgggtcagt atgaaagcca taccgcattt 1500
acactgcctg gtctgtatcg tgttgttcat ggtattgatg tgttcgaccc gaaatttaac 1560
attgttagtc cgggtgcaga tcagaccatc tattttccgc ataccgaaac cagccgtcgc 1620
ctgaccagct ttcatccgga aattgaggaa ctgctgtata gcagcgttga aaacgaagaa 1680
catatttgcg ttctgaaaga tcgtagcaaa ccgatcattt ttaccatggc acgcctggat 1740
cgtgttaaaa acattaccgg tctggttgaa tggtatggca aaaatgcaaa actgcgcgaa 1800
ctggttaatc tggttgtggt tgccggtgat cgtcgtaaag aaagtaaaga tctggaagaa 1860
aaagccgaaa tgaagaaaat gtatggcctg atcgaaacct ataaactgaa tggccagttt 1920
cgttggatta gcagccagat gaatcgtgtt cgtaatggtg aactgtatcg cgttatttgt 1980
gatacccgtg gtgcctttgt tcagcctgcc gtttatgaag cctttggtct gaccgttgtg 2040
gaagcaatga cctgcggtct gccgaccttt gcaacctgta atggtggtcc ggcagaaatt 2100
attgtgcatg gtaaatccgg ttttcacatc gatccgtatc atggtgatcg tgcagcagac 2160
ctgctggttg atttttttga aaaatgtaaa ctggatccga cgcactggga taaaatcagc 2220
aaagccggtc tgcagcgcat tgaagagaaa tatacctggc agatttatag ccagcgtctg 2280
ctgaccctga caggtgttta tggtttttgg aaacatgtga gcaatctgga tcgtcgtgaa 2340
tcacgtcgtt acctggaaat gttttatgcc ctgaaatatc gcaaactggc agaaagcgtt 2400
ccgctggcag cagaataa 2418
<210> 211
<211> 339
<212> PRT
<213> B. subtillis
<400> 211
Met Ala Ile Leu Val Thr Gly Gly Ala Gly Tyr Ile Gly Ser His Thr
1 5 10 15
Cys Val Glu Leu Leu Asn Ser Gly Tyr Glu Ile Val Val Leu Asp Asn
20 25 30
Leu Ser Asn Ser Ser Ala Glu Ala Leu Asn Arg Val Lys Glu Ile Thr
35 40 45
Gly Lys Asp Leu Thr Phe Tyr Glu Ala Asp Leu Leu Asp Arg Glu Ala
50 55 60
Val Asp Ser Val Phe Ala Glu Asn Glu Ile Glu Ala Val Ile His Phe
65 70 75 80
Ala Gly Leu Lys Ala Val Gly Glu Ser Val Ala Ile Pro Leu Lys Tyr
85 90 95
Tyr His Asn Asn Leu Thr Gly Thr Phe Ile Leu Cys Glu Ala Met Glu
100 105 110
Lys Tyr Gly Val Lys Lys Ile Val Phe Ser Ser Ser Ala Thr Val Tyr
115 120 125
Gly Val Pro Glu Thr Ser Pro Ile Thr Glu Asp Phe Pro Leu Gly Ala
130 135 140
Thr Asn Pro Tyr Gly Gln Thr Lys Leu Met Leu Glu Gln Ile Leu Arg
145 150 155 160
Asp Leu His Thr Ala Asp Asn Glu Trp Ser Val Ala Leu Leu Arg Tyr
165 170 175
Phe Asn Pro Phe Gly Ala His Pro Ser Gly Arg Ile Gly Glu Asp Pro
180 185 190
Asn Gly Ile Pro Asn Asn Leu Met Pro Tyr Val Ala Gln Val Ala Val
195 200 205
Gly Lys Leu Glu Gln Leu Ser Val Phe Gly Asn Asp Tyr Pro Thr Lys
210 215 220
Asp Gly Thr Gly Val Arg Asp Tyr Ile His Val Val Asp Leu Ala Glu
225 230 235 240
Gly His Val Lys Ala Leu Glu Lys Val Leu Asn Ser Thr Gly Ala Asp
245 250 255
Ala Tyr Asn Leu Gly Thr Gly Thr Gly Tyr Ser Val Leu Glu Met Val
260 265 270
Lys Ala Phe Glu Lys Val Ser Gly Lys Glu Val Pro Tyr Arg Phe Ala
275 280 285
Asp Arg Arg Pro Gly Asp Ile Ala Thr Cys Phe Ala Asp Pro Ala Lys
290 295 300
Ala Lys Arg Glu Leu Gly Trp Glu Ala Lys Arg Gly Leu Glu Glu Met
305 310 315 320
Cys Ala Asp Ser Trp Arg Trp Gln Ser Ser Asn Val Asn Gly Tyr Lys
325 330 335
Ser Ala Glu
<210> 212
<211> 1020
<212> DNA
<213> B. subtillis
<400> 212
atggcaatac ttgttactgg cggtgccggt tacattggca gccacacatg tgttgaacta 60
ttgaacagcg gctacgagat tgttgttctt gataatctgt ccaacagttc agctgaagcg 120
ctgaaccgtg tcaaggagat tacaggaaaa gatttaacgt tctacgaagc ggatttattg 180
gaccgggaag cggtagattc cgtttttgct gaaaatgaaa tcgaagctgt gattcatttt 240
gcagggttaa aagcagtcgg cgaatctgtg gcgattcccc tcaaatatta tcataacaat 300
ttgacaggaa cgtttatttt atgcgaggcc atggagaaat acggcgtcaa gaaaatcgta 360
ttcagttcat ctgcgacagt atacggcgtt ccggaaacat cgccgattac ggaagacttt 420
ccattaggcg cgacaaatcc ttatgggcag acgaagctca tgcttgaaca aatattgcgt 480
gatttgcata cagccgacaa tgagtggagc gttgcgctgc ttcgttactt taacccgttc 540
ggcgcgcatc caagcggacg gatcggtgaa gacccgaacg gaatcccaaa taaccttatg 600
ccgtatgtgg cacaggtagc agtcgggaag ctcgagcaat taagcgtatt cggaaatgac 660
tatccgacaa aagacgggac aggcgtacgc gattatattc acgtcgttga tctcgcagaa 720
ggccacgtca aggcgctgga aaaagtattg aactctacag gagccgatgc atacaacctt 780
ggaacaggca caggctacag cgtgctggaa atggtcaaag cctttgaaaa agtgtcaggg 840
aaagaggttc cataccgttt tgcggaccgc cgtccgggag acatcgccac atgctttgca 900
gatcctgcga aagccaagcg agaactaggc tgggaagcga aacgcggcct tgaggaaatg 960
tgtgctgatt cctggagatg gcagtcttct aatgtgaatg ggtataagag tgcggaataa 1020
<210> 213
<211> 342
<212> PRT
<213> 拟南芥
<400> 213
Met Ala Ala Thr Ser Glu Lys Gln Asn Thr Thr Lys Pro Pro Pro Ser
1 5 10 15
Pro Ser Pro Leu Arg Asn Ser Lys Phe Cys Gln Pro Asn Met Arg Ile
20 25 30
Leu Ile Ser Gly Gly Ala Gly Phe Ile Gly Ser His Leu Val Asp Lys
35 40 45
Leu Met Glu Asn Glu Lys Asn Glu Val Val Val Ala Asp Asn Tyr Phe
50 55 60
Thr Gly Ser Lys Glu Asn Leu Lys Lys Trp Ile Gly His Pro Arg Phe
65 70 75 80
Glu Leu Ile Arg His Asp Val Thr Glu Pro Leu Leu Ile Glu Val Asp
85 90 95
Arg Ile Tyr His Leu Ala Cys Pro Ala Ser Pro Ile Phe Tyr Lys Tyr
100 105 110
Asn Pro Val Lys Thr Ile Lys Thr Asn Val Ile Gly Thr Leu Asn Met
115 120 125
Leu Gly Leu Ala Lys Arg Val Gly Ala Arg Ile Leu Leu Thr Ser Thr
130 135 140
Ser Glu Val Tyr Gly Asp Pro Leu Ile His Pro Gln Pro Glu Ser Tyr
145 150 155 160
Trp Gly Asn Val Asn Pro Ile Gly Val Arg Ser Cys Tyr Asp Glu Gly
165 170 175
Lys Arg Val Ala Glu Thr Leu Met Phe Asp Tyr His Arg Gln His Gly
180 185 190
Ile Glu Ile Arg Ile Ala Arg Ile Phe Asn Thr Tyr Gly Pro Arg Met
195 200 205
Asn Ile Asp Asp Gly Arg Val Val Ser Asn Phe Ile Ala Gln Ala Leu
210 215 220
Arg Gly Glu Ala Leu Thr Val Gln Lys Pro Gly Thr Gln Thr Arg Ser
225 230 235 240
Phe Cys Tyr Val Ser Asp Met Val Asp Gly Leu Ile Arg Leu Met Glu
245 250 255
Gly Asn Asp Thr Gly Pro Ile Asn Ile Gly Asn Pro Gly Glu Phe Thr
260 265 270
Met Val Glu Leu Ala Glu Thr Val Lys Glu Leu Ile Asn Pro Ser Ile
275 280 285
Glu Ile Lys Met Val Glu Asn Thr Pro Asp Asp Pro Arg Gln Arg Lys
290 295 300
Pro Asp Ile Ser Lys Ala Lys Glu Val Leu Gly Trp Glu Pro Lys Val
305 310 315 320
Lys Leu Arg Glu Gly Leu Pro Leu Met Glu Glu Asp Phe Arg Leu Arg
325 330 335
Leu Asn Val Pro Arg Asn
340
<210> 214
<211> 1029
<212> DNA
<213> 拟南芥
<400> 214
atggcagcta caagtgagaa acagaacacc acaaagcctc ctccttctcc ttctcctctc 60
cgcaattcca agttttgtca gcccaatatg aggatcttga tctctggagg agctggcttc 120
attggttctc acttggttga taagcttatg gaaaatgaga agaatgaggt ggttgttgct 180
gataactatt tcactggctc aaaagaaaac ctcaagaagt ggatcggtca ccccaggttt 240
gaacttattc gtcacgatgt taccgagcct ttgttgatcg aggttgatcg gatttaccat 300
cttgcttgtc ctgcctctcc tatcttctac aaatacaacc ctgttaagac aatcaagacc 360
aatgtgattg gtacactcaa catgctcggt cttgccaagc gtgttggagc aagaatttta 420
ctaacctcaa cctctgaagt gtatggagat cctctcatcc accctcaacc agagagctac 480
tggggaaatg tcaaccctat tggggttcgg agttgctatg acgaaggcaa gcgggtagcc 540
gaaaccttga tgtttgacta ccacagacaa catggcattg aaatccgcat tgctagaatc 600
ttcaacacat atggtcctcg aatgaacatc gatgatgggc gtgttgtgag caacttcatt 660
gctcaagcac tccggggtga ggcattgaca gttcagaaac cggggacaca gacccgcagt 720
ttctgttatg tctccgacat ggtggatgga cttatccgtc ttatggaagg caatgatact 780
ggccctatca acatcggtaa cccaggtgag ttcacaatgg tggaactggc tgagacggtt 840
aaggagctta ttaacccaag catagagata aagatggtgg agaacacacc agatgatcca 900
agacagagga aaccagacat tagtaaagcc aaagaagtgt tgggttggga gccaaaggtg 960
aagctcagag aaggacttcc tctcatggaa gaagatttcc gactaaggct taacgtccca 1020
agaaactaa 1029
<210> 215
<211> 297
<212> PRT
<213> 拟南芥
<400> 215
Thr Pro Lys Asn Gly Asp Ser Gly Asp Lys Ala Ser Leu Lys Phe Leu
1 5 10 15
Ile Tyr Gly Lys Thr Gly Trp Leu Gly Gly Leu Leu Gly Lys Leu Cys
20 25 30
Glu Lys Gln Gly Ile Thr Tyr Glu Tyr Gly Lys Gly Arg Leu Glu Asp
35 40 45
Arg Ala Ser Leu Val Ala Asp Ile Arg Ser Ile Lys Pro Thr His Val
50 55 60
Phe Asn Ala Ala Gly Leu Thr Gly Arg Pro Asn Val Asp Trp Cys Glu
65 70 75 80
Ser His Lys Pro Glu Thr Ile Arg Val Asn Val Ala Gly Thr Leu Thr
85 90 95
Leu Ala Asp Val Cys Arg Glu Asn Asp Leu Leu Met Met Asn Phe Ala
100 105 110
Thr Gly Cys Ile Phe Glu Tyr Asp Ala Thr His Pro Glu Gly Ser Gly
115 120 125
Ile Gly Phe Lys Glu Glu Asp Lys Pro Asn Phe Phe Gly Ser Phe Tyr
130 135 140
Ser Lys Thr Lys Ala Met Val Glu Glu Leu Leu Arg Glu Phe Asp Asn
145 150 155 160
Val Cys Thr Leu Arg Val Arg Met Pro Ile Ser Ser Asp Leu Asn Asn
165 170 175
Pro Arg Asn Phe Ile Thr Lys Ile Ser Arg Tyr Asn Lys Val Val Asp
180 185 190
Ile Pro Asn Ser Met Thr Val Leu Asp Glu Leu Leu Pro Ile Ser Ile
195 200 205
Glu Met Ala Lys Arg Asn Leu Arg Gly Ile Trp Asn Phe Thr Asn Pro
210 215 220
Gly Val Val Ser His Asn Glu Ile Leu Glu Met Tyr Lys Asn Tyr Ile
225 230 235 240
Glu Pro Gly Phe Lys Trp Ser Asn Phe Thr Val Glu Glu Gln Ala Lys
245 250 255
Val Ile Val Ala Ala Arg Ser Asn Asn Glu Met Asp Gly Ser Lys Leu
260 265 270
Ser Lys Glu Phe Pro Glu Met Leu Ser Ile Lys Glu Ser Leu Leu Lys
275 280 285
Tyr Val Phe Glu Pro Asn Lys Arg Thr
290 295
<210> 216
<211> 894
<212> DNA
<213> 拟南芥
<400> 216
acacctaaga atggtgattc tggtgacaaa gcttcgttga agtttttgat ctatggtaag 60
actggttggc ttggtggtct tctagggaaa ctatgtgaga agcaagggat tacatatgag 120
tatgggaaag gacgtctgga ggatagagct tctcttgtgg cggatattcg tagcatcaaa 180
cctactcatg tgtttaatgc tgctggttta actggcagac ccaacgttga ctggtgtgaa 240
tctcacaaac cagagaccat tcgtgtaaat gtcgcaggta ctttgactct agctgatgtt 300
tgcagagaga atgatctctt gatgatgaac ttcgccaccg gttgcatctt tgagtatgac 360
gctacacatc ctgagggttc gggtataggt ttcaaggaag aagacaagcc aaatttcttt 420
ggttctttct actcgaaaac caaagccatg gttgaggagc tcttgagaga atttgacaat 480
gtatgtacct tgagagtccg gatgccaatc tcctcagacc taaacaaccc gagaaacttc 540
atcacgaaga tctcgcgcta caacaaagtg gtggacatcc cgaacagcat gaccgtacta 600
gacgagcttc tcccaatctc tatcgagatg gcgaagagaa acctaagagg catatggaat 660
ttcaccaacc caggggtggt gagccacaac gagatattgg agatgtacaa gaattacatc 720
gagccaggtt ttaaatggtc caacttcaca gtggaagaac aagcaaaggt cattgttgct 780
gctcgaagca acaacgaaat ggatggatct aaactaagca aggagttccc agagatgctc 840
tccatcaaag agtcactgct caaatacgtc tttgaaccaa acaagagaac ctaa 894
<210> 217
<211> 370
<212> PRT
<213> 拟南芥
<400> 217
Met Asp Asp Thr Thr Tyr Lys Pro Lys Asn Ile Leu Ile Thr Gly Ala
1 5 10 15
Ala Gly Phe Ile Ala Ser His Val Ala Asn Arg Leu Ile Arg Asn Tyr
20 25 30
Pro Asp Tyr Lys Ile Val Val Leu Asp Lys Leu Asp Tyr Cys Ser Asp
35 40 45
Leu Lys Asn Leu Asp Pro Ser Phe Ser Ser Pro Asn Phe Lys Phe Val
50 55 60
Lys Gly Asp Ile Ala Ser Asp Asp Leu Val Asn Tyr Leu Leu Ile Thr
65 70 75 80
Glu Asn Ile Asp Thr Ile Met His Phe Ala Ala Gln Thr His Val Asp
85 90 95
Asn Ser Phe Gly Asn Ser Phe Glu Phe Thr Lys Asn Asn Ile Tyr Gly
100 105 110
Thr His Val Leu Leu Glu Ala Cys Lys Val Thr Gly Gln Ile Arg Arg
115 120 125
Phe Ile His Val Ser Thr Asp Glu Val Tyr Gly Glu Thr Asp Glu Asp
130 135 140
Ala Ala Val Gly Asn His Glu Ala Ser Gln Leu Leu Pro Thr Asn Pro
145 150 155 160
Tyr Ser Ala Thr Lys Ala Gly Ala Glu Met Leu Val Met Ala Tyr Gly
165 170 175
Arg Ser Tyr Gly Leu Pro Val Ile Thr Thr Arg Gly Asn Asn Val Tyr
180 185 190
Gly Pro Asn Gln Phe Pro Glu Lys Met Ile Pro Lys Phe Ile Leu Leu
195 200 205
Ala Met Ser Gly Lys Pro Leu Pro Ile His Gly Asp Gly Ser Asn Val
210 215 220
Arg Ser Tyr Leu Tyr Cys Glu Asp Val Ala Glu Ala Phe Glu Val Val
225 230 235 240
Leu His Lys Gly Glu Ile Gly His Val Tyr Asn Val Gly Thr Lys Arg
245 250 255
Glu Arg Arg Val Ile Asp Val Ala Arg Asp Ile Cys Lys Leu Phe Gly
260 265 270
Lys Asp Pro Glu Ser Ser Ile Gln Phe Val Glu Asn Arg Pro Phe Asn
275 280 285
Asp Gln Arg Tyr Phe Leu Asp Asp Gln Lys Leu Lys Lys Leu Gly Trp
290 295 300
Gln Glu Arg Thr Asn Trp Glu Asp Gly Leu Lys Lys Thr Met Asp Trp
305 310 315 320
Tyr Thr Gln Asn Pro Glu Trp Trp Gly Asp Val Ser Gly Ala Leu Leu
325 330 335
Pro His Pro Arg Met Leu Met Met Pro Gly Gly Arg Leu Ser Asp Gly
340 345 350
Ser Ser Glu Lys Lys Asp Val Ser Ser Asn Thr Val Gln Thr Phe Thr
355 360 365
Val Val
370
<210> 218
<211> 1113
<212> DNA
<213> 拟南芥
<400> 218
atggatgata ctacgtataa gccaaagaac attctcatta ctggagctgc tggatttatt 60
gcttctcatg ttgccaacag attaatccgt aactatcctg attacaagat cgttgttctt 120
gacaagcttg attactgttc agatctgaag aatcttgatc cttctttttc ttcaccaaat 180
ttcaagtttg tcaaaggaga tatcgcgagt gatgatctcg ttaactacct tctcatcact 240
gaaaacattg atacgataat gcattttgct gctcaaactc atgttgataa ctcttttggt 300
aatagctttg agtttaccaa gaacaatatt tatggtactc atgttctttt ggaagcctgt 360
aaagttacag gacagatcag gaggtttatc catgtgagta ccgatgaagt ctatggagaa 420
accgatgagg atgctgctgt aggaaaccat gaagcttctc agctgttacc gacgaatcct 480
tactctgcaa ctaaggctgg tgctgagatg cttgtgatgg cttatggtag atcatatgga 540
ttgcctgtta ttacgactcg cgggaacaat gtttatgggc ctaaccagtt tcctgaaaaa 600
atgattccta agttcatctt gttggctatg agtgggaagc cgcttcccat ccatggagat 660
ggatctaatg tccggagtta cttgtactgc gaagacgttg ctgaggcttt tgaggttgtt 720
cttcacaaag gagaaatcgg tcatgtctac aatgtcggca caaaaagaga aaggagagtg 780
atcgatgtgg ctagagacat ctgcaaactt ttcgggaaag accctgagtc aagcattcag 840
tttgtggaga accggccctt taatgatcaa aggtacttcc ttgatgatca gaagctgaag 900
aaattggggt ggcaagagcg aacaaattgg gaagatggat tgaagaagac aatggactgg 960
tacactcaga atcctgagtg gtggggtgat gtttctggag ctttgcttcc tcatccgaga 1020
atgcttatga tgcccggtgg aagactttct gatggatcta gtgagaagaa agacgtttca 1080
agcaacacgg tccagacatt tacggttgta taa 1113
<210> 219
<211> 667
<212> PRT
<213> 拟南芥
<400> 219
Met Asp Asp Thr Thr Tyr Lys Pro Lys Asn Ile Leu Ile Thr Gly Ala
1 5 10 15
Ala Gly Phe Ile Ala Ser His Val Ala Asn Arg Leu Ile Arg Asn Tyr
20 25 30
Pro Asp Tyr Lys Ile Val Val Leu Asp Lys Leu Asp Tyr Cys Ser Asp
35 40 45
Leu Lys Asn Leu Asp Pro Ser Phe Ser Ser Pro Asn Phe Lys Phe Val
50 55 60
Lys Gly Asp Ile Ala Ser Asp Asp Leu Val Asn Tyr Leu Leu Ile Thr
65 70 75 80
Glu Asn Ile Asp Thr Ile Met His Phe Ala Ala Gln Thr His Val Asp
85 90 95
Asn Ser Phe Gly Asn Ser Phe Glu Phe Thr Lys Asn Asn Ile Tyr Gly
100 105 110
Thr His Val Leu Leu Glu Ala Cys Lys Val Thr Gly Gln Ile Arg Arg
115 120 125
Phe Ile His Val Ser Thr Asp Glu Val Tyr Gly Glu Thr Asp Glu Asp
130 135 140
Ala Ala Val Gly Asn His Glu Ala Ser Gln Leu Leu Pro Thr Asn Pro
145 150 155 160
Tyr Ser Ala Thr Lys Ala Gly Ala Glu Met Leu Val Met Ala Tyr Gly
165 170 175
Arg Ser Tyr Gly Leu Pro Val Ile Thr Thr Arg Gly Asn Asn Val Tyr
180 185 190
Gly Pro Asn Gln Phe Pro Glu Lys Met Ile Pro Lys Phe Ile Leu Leu
195 200 205
Ala Met Ser Gly Lys Pro Leu Pro Ile His Gly Asp Gly Ser Asn Val
210 215 220
Arg Ser Tyr Leu Tyr Cys Glu Asp Val Ala Glu Ala Phe Glu Val Val
225 230 235 240
Leu His Lys Gly Glu Ile Gly His Val Tyr Asn Val Gly Thr Lys Arg
245 250 255
Glu Arg Arg Val Ile Asp Val Ala Arg Asp Ile Cys Lys Leu Phe Gly
260 265 270
Lys Asp Pro Glu Ser Ser Ile Gln Phe Val Glu Asn Arg Pro Phe Asn
275 280 285
Asp Gln Arg Tyr Phe Leu Asp Asp Gln Lys Leu Lys Lys Leu Gly Trp
290 295 300
Gln Glu Arg Thr Asn Trp Glu Asp Gly Leu Lys Lys Thr Met Asp Trp
305 310 315 320
Tyr Thr Gln Asn Pro Glu Trp Trp Gly Asp Val Ser Gly Ala Leu Leu
325 330 335
Pro His Pro Arg Met Leu Met Met Pro Gly Gly Arg Leu Ser Asp Gly
340 345 350
Ser Ser Glu Lys Lys Asp Val Ser Ser Asn Thr Val Gln Thr Phe Thr
355 360 365
Val Val Thr Pro Lys Asn Gly Asp Ser Gly Asp Lys Ala Ser Leu Lys
370 375 380
Phe Leu Ile Tyr Gly Lys Thr Gly Trp Leu Gly Gly Leu Leu Gly Lys
385 390 395 400
Leu Cys Glu Lys Gln Gly Ile Thr Tyr Glu Tyr Gly Lys Gly Arg Leu
405 410 415
Glu Asp Arg Ala Ser Leu Val Ala Asp Ile Arg Ser Ile Lys Pro Thr
420 425 430
His Val Phe Asn Ala Ala Gly Leu Thr Gly Arg Pro Asn Val Asp Trp
435 440 445
Cys Glu Ser His Lys Pro Glu Thr Ile Arg Val Asn Val Ala Gly Thr
450 455 460
Leu Thr Leu Ala Asp Val Cys Arg Glu Asn Asp Leu Leu Met Met Asn
465 470 475 480
Phe Ala Thr Gly Cys Ile Phe Glu Tyr Asp Ala Thr His Pro Glu Gly
485 490 495
Ser Gly Ile Gly Phe Lys Glu Glu Asp Lys Pro Asn Phe Phe Gly Ser
500 505 510
Phe Tyr Ser Lys Thr Lys Ala Met Val Glu Glu Leu Leu Arg Glu Phe
515 520 525
Asp Asn Val Cys Thr Leu Arg Val Arg Met Pro Ile Ser Ser Asp Leu
530 535 540
Asn Asn Pro Arg Asn Phe Ile Thr Lys Ile Ser Arg Tyr Asn Lys Val
545 550 555 560
Val Asp Ile Pro Asn Ser Met Thr Val Leu Asp Glu Leu Leu Pro Ile
565 570 575
Ser Ile Glu Met Ala Lys Arg Asn Leu Arg Gly Ile Trp Asn Phe Thr
580 585 590
Asn Pro Gly Val Val Ser His Asn Glu Ile Leu Glu Met Tyr Lys Asn
595 600 605
Tyr Ile Glu Pro Gly Phe Lys Trp Ser Asn Phe Thr Val Glu Glu Gln
610 615 620
Ala Lys Val Ile Val Ala Ala Arg Ser Asn Asn Glu Met Asp Gly Ser
625 630 635 640
Lys Leu Ser Lys Glu Phe Pro Glu Met Leu Ser Ile Lys Glu Ser Leu
645 650 655
Leu Lys Tyr Val Phe Glu Pro Asn Lys Arg Thr
660 665
<210> 220
<211> 2004
<212> DNA
<213> 拟南芥
<400> 220
atggatgata ctacgtataa gccaaagaac attctcatta ctggagctgc tggatttatt 60
gcttctcatg ttgccaacag attaatccgt aactatcctg attacaagat cgttgttctt 120
gacaagcttg attactgttc agatctgaag aatcttgatc cttctttttc ttcaccaaat 180
ttcaagtttg tcaaaggaga tatcgcgagt gatgatctcg ttaactacct tctcatcact 240
gaaaacattg atacgataat gcattttgct gctcaaactc atgttgataa ctcttttggt 300
aatagctttg agtttaccaa gaacaatatt tatggtactc atgttctttt ggaagcctgt 360
aaagttacag gacagatcag gaggtttatc catgtgagta ccgatgaagt ctatggagaa 420
accgatgagg atgctgctgt aggaaaccat gaagcttctc agctgttacc gacgaatcct 480
tactctgcaa ctaaggctgg tgctgagatg cttgtgatgg cttatggtag atcatatgga 540
ttgcctgtta ttacgactcg cgggaacaat gtttatgggc ctaaccagtt tcctgaaaaa 600
atgattccta agttcatctt gttggctatg agtgggaagc cgcttcccat ccatggagat 660
ggatctaatg tccggagtta cttgtactgc gaagacgttg ctgaggcttt tgaggttgtt 720
cttcacaaag gagaaatcgg tcatgtctac aatgtcggca caaaaagaga aaggagagtg 780
atcgatgtgg ctagagacat ctgcaaactt ttcgggaaag accctgagtc aagcattcag 840
tttgtggaga accggccctt taatgatcaa aggtacttcc ttgatgatca gaagctgaag 900
aaattggggt ggcaagagcg aacaaattgg gaagatggat tgaagaagac aatggactgg 960
tacactcaga atcctgagtg gtggggtgat gtttctggag ctttgcttcc tcatccgaga 1020
atgcttatga tgcccggtgg aagactttct gatggatcta gtgagaagaa agacgtttca 1080
agcaacacgg tccagacatt tacggttgta acacctaaga atggtgattc tggtgacaaa 1140
gcttcgttga agtttttgat ctatggtaag actggttggc ttggtggtct tctagggaaa 1200
ctatgtgaga agcaagggat tacatatgag tatgggaaag gacgtctgga ggatagagct 1260
tctcttgtgg cggatattcg tagcatcaaa cctactcatg tgtttaatgc tgctggttta 1320
actggcagac ccaacgttga ctggtgtgaa tctcacaaac cagagaccat tcgtgtaaat 1380
gtcgcaggta ctttgactct agctgatgtt tgcagagaga atgatctctt gatgatgaac 1440
ttcgccaccg gttgcatctt tgagtatgac gctacacatc ctgagggttc gggtataggt 1500
ttcaaggaag aagacaagcc aaatttcttt ggttctttct actcgaaaac caaagccatg 1560
gttgaggagc tcttgagaga atttgacaat gtatgtacct tgagagtccg gatgccaatc 1620
tcctcagacc taaacaaccc gagaaacttc atcacgaaga tctcgcgcta caacaaagtg 1680
gtggacatcc cgaacagcat gaccgtacta gacgagcttc tcccaatctc tatcgagatg 1740
gcgaagagaa acctaagagg catatggaat ttcaccaacc caggggtggt gagccacaac 1800
gagatattgg agatgtacaa gaattacatc gagccaggtt ttaaatggtc caacttcaca 1860
gtggaagaac aagcaaaggt cattgttgct gctcgaagca acaacgaaat ggatggatct 1920
aaactaagca aggagttccc agagatgctc tccatcaaag agtcactgct caaatacgtc 1980
tttgaaccaa acaagagaac ctaa 2004
<210> 221
<211> 481
<212> PRT
<213> 拟南芥
<400> 221
Met Val Lys Ile Cys Cys Ile Gly Ala Gly Tyr Val Gly Gly Pro Thr
1 5 10 15
Met Ala Val Met Ala Leu Lys Cys Pro Glu Ile Glu Val Val Val Val
20 25 30
Asp Ile Ser Glu Pro Arg Ile Asn Ala Trp Asn Ser Asp Arg Leu Pro
35 40 45
Ile Tyr Glu Pro Gly Leu Glu Asp Val Val Lys Gln Cys Arg Gly Lys
50 55 60
Asn Leu Phe Phe Ser Thr Asp Val Glu Lys His Val Phe Glu Ser Asp
65 70 75 80
Ile Val Phe Val Ser Val Asn Thr Pro Thr Lys Thr Gln Gly Leu Gly
85 90 95
Ala Gly Lys Ala Ala Asp Leu Thr Tyr Trp Glu Ser Ala Ala Arg Met
100 105 110
Ile Ala Asp Val Ser Lys Ser Ser Lys Ile Val Val Glu Lys Ser Thr
115 120 125
Val Pro Val Arg Thr Ala Glu Ala Ile Glu Lys Ile Leu Thr His Asn
130 135 140
Ser Lys Gly Ile Glu Phe Gln Ile Leu Ser Asn Pro Glu Phe Leu Ala
145 150 155 160
Glu Gly Thr Ala Ile Lys Asp Leu Tyr Asn Pro Asp Arg Val Leu Ile
165 170 175
Gly Gly Arg Asp Thr Ala Ala Gly Gln Lys Ala Ile Lys Ala Leu Arg
180 185 190
Asp Val Tyr Ala His Trp Val Pro Val Glu Gln Ile Ile Cys Thr Asn
195 200 205
Leu Trp Ser Ala Glu Leu Ser Lys Leu Ala Ala Asn Ala Phe Leu Ala
210 215 220
Gln Arg Ile Ser Ser Val Asn Ala Met Ser Ala Leu Cys Glu Ala Thr
225 230 235 240
Gly Ala Asp Val Thr Gln Val Ala His Ala Val Gly Thr Asp Thr Arg
245 250 255
Ile Gly Pro Lys Phe Leu Asn Ala Ser Val Gly Phe Gly Gly Ser Cys
260 265 270
Phe Gln Lys Asp Ile Leu Asn Leu Ile Tyr Ile Cys Glu Cys Asn Gly
275 280 285
Leu Pro Glu Ala Ala Asn Tyr Trp Lys Gln Val Val Lys Val Asn Asp
290 295 300
Tyr Gln Lys Ile Arg Phe Ala Asn Arg Val Val Ser Ser Met Phe Asn
305 310 315 320
Thr Val Ser Gly Lys Lys Ile Ala Ile Leu Gly Phe Ala Phe Lys Lys
325 330 335
Asp Thr Gly Asp Thr Arg Glu Thr Pro Ala Ile Asp Val Cys Asn Arg
340 345 350
Leu Val Ala Asp Lys Ala Lys Leu Ser Ile Tyr Asp Pro Gln Val Leu
355 360 365
Glu Glu Gln Ile Arg Arg Asp Leu Ser Met Ala Arg Phe Asp Trp Asp
370 375 380
His Pro Val Pro Leu Gln Gln Ile Lys Ala Glu Gly Ile Ser Glu Gln
385 390 395 400
Val Asn Val Val Ser Asp Ala Tyr Glu Ala Thr Lys Asp Ala His Gly
405 410 415
Leu Cys Val Leu Thr Glu Trp Asp Glu Phe Lys Ser Leu Asp Phe Lys
420 425 430
Lys Ile Phe Asp Asn Met Gln Lys Pro Ala Phe Val Phe Asp Gly Arg
435 440 445
Asn Val Val Asp Ala Val Lys Leu Arg Glu Ile Gly Phe Ile Val Tyr
450 455 460
Ser Ile Gly Lys Pro Leu Asp Ser Trp Leu Lys Asp Met Pro Ala Val
465 470 475 480
Ala
<210> 222
<211> 1446
<212> DNA
<213> 拟南芥
<400> 222
atggtgaaaa tttgttgtat tggcgcaggt tatgttggtg gtccgaccat ggcagttatg 60
gcactgaaat gtccggaaat tgaagttgtt gttgtggata ttagcgaacc gcgtattaat 120
gcatggaata gcgatcgtct gccgatttat gaacctggtc tggaagatgt tgttaaacag 180
tgtcgtggta aaaacctgtt ttttagcacc gatgtggaaa agcatgtgtt tgaaagcgat 240
attgttttcg tgagcgttaa taccccgacc aaaacacaag gtttaggtgc aggtaaagca 300
gccgatctga cctattggga aagcgcagca cgtatgattg cagatgttag caaaagcagc 360
aaaatcgtgg ttgaaaaaag caccgttccg gttcgtaccg cagaagcaat tgaaaaaatt 420
ctgacccata acagcaaagg catcgaattt cagattctga gcaatccgga atttctggca 480
gaaggcaccg caattaaaga tctgtataat ccggatcgtg ttctgattgg tggtcgtgat 540
accgcagcag gtcagaaagc cattaaagca ctgcgtgatg tttatgcaca ttgggttcca 600
gttgagcaga ttatttgtac caatctgtgg tcagcagaac tgagcaaact ggcagcaaat 660
gcctttctgg cacagcgtat tagcagcgtt aatgcaatga gcgcactgtg tgaagcaacc 720
ggtgccgatg ttacccaggt tgcacatgca gttggtacag atacccgtat tggtccgaaa 780
tttctgaatg caagcgttgg ttttggtggt agctgttttc agaaagatat tctgaacctg 840
atctacatct gcgaatgtaa tggtctgccg gaagcagcca attattggaa acaggttgtt 900
aaagtgaacg attaccagaa aattcgcttt gccaatcgtg ttgttagcag catgtttaat 960
accgtgagcg gcaaaaaaat cgccattctg ggttttgcct tcaaaaaaga taccggtgat 1020
acccgtgaaa caccggcaat tgatgtttgt aatcgtctgg ttgcagataa agccaaactg 1080
agcatttatg atccgcaggt tctggaagaa caaattcgtc gtgatctgag catggcacgt 1140
tttgattggg atcatccggt tccgctgcag cagattaaag cagaaggtat ttcagaacag 1200
gtgaacgttg ttagtgatgc atatgaagcc accaaagatg cacatggtct gtgtgttctg 1260
accgaatggg atgaattcaa aagcctggat ttcaaaaaga tcttcgataa catgcagaaa 1320
ccggcatttg tttttgatgg tcgtaatgtt gttgatgccg ttaaactgcg tgaaatcggc 1380
tttattgttt acagcattgg taaaccgctg gatagctggc tgaaagatat gcctgcagtt 1440
gcataa 1446
<210> 223
<211> 419
<212> PRT
<213> 拟南芥
<400> 223
Met Phe Ser Phe Gly Arg Ala Arg Ser Gln Gly Arg Gln Asn Arg Ser
1 5 10 15
Met Ser Leu Gly Gly Leu Asp Tyr Ala Asp Pro Lys Lys Lys Asn Asn
20 25 30
Tyr Leu Gly Lys Ile Leu Leu Thr Ala Ser Leu Thr Ala Leu Cys Ile
35 40 45
Phe Met Leu Lys Gln Ser Pro Thr Phe Asn Thr Pro Ser Val Phe Ser
50 55 60
Arg His Glu Pro Gly Val Thr His Val Leu Val Thr Gly Gly Ala Gly
65 70 75 80
Tyr Ile Gly Ser His Ala Ala Leu Arg Leu Leu Lys Glu Ser Tyr Arg
85 90 95
Val Thr Ile Val Asp Asn Leu Ser Arg Gly Asn Leu Ala Ala Val Arg
100 105 110
Ile Leu Gln Glu Leu Phe Pro Glu Pro Gly Arg Leu Gln Phe Ile Tyr
115 120 125
Ala Asp Leu Gly Asp Ala Lys Ala Val Asn Lys Ile Phe Thr Glu Asn
130 135 140
Ala Phe Asp Ala Val Met His Phe Ala Ala Val Ala Tyr Val Gly Glu
145 150 155 160
Ser Thr Gln Phe Pro Leu Lys Tyr Tyr His Asn Ile Thr Ser Asn Thr
165 170 175
Leu Val Val Leu Glu Thr Met Ala Ala His Gly Val Lys Thr Leu Ile
180 185 190
Tyr Ser Ser Thr Cys Ala Thr Tyr Gly Glu Pro Asp Ile Met Pro Ile
195 200 205
Thr Glu Glu Thr Pro Gln Val Pro Ile Asn Pro Tyr Gly Lys Ala Lys
210 215 220
Lys Met Ala Glu Asp Ile Ile Leu Asp Phe Ser Lys Asn Ser Asp Met
225 230 235 240
Ala Val Met Ile Leu Arg Tyr Phe Asn Val Ile Gly Ser Asp Pro Glu
245 250 255
Gly Arg Leu Gly Glu Ala Pro Arg Pro Glu Leu Arg Glu His Gly Arg
260 265 270
Ile Ser Gly Ala Cys Phe Asp Ala Ala Arg Gly Ile Met Pro Gly Leu
275 280 285
Gln Ile Lys Gly Thr Asp Tyr Lys Thr Ala Asp Gly Thr Cys Val Arg
290 295 300
Asp Tyr Ile Asp Val Thr Asp Leu Val Asp Ala His Val Lys Ala Leu
305 310 315 320
Gln Lys Ala Lys Pro Arg Lys Val Gly Ile Tyr Asn Val Gly Thr Gly
325 330 335
Lys Gly Ser Ser Val Lys Glu Phe Val Glu Ala Cys Lys Lys Ala Thr
340 345 350
Gly Val Glu Ile Lys Ile Asp Tyr Leu Pro Arg Arg Ala Gly Asp Tyr
355 360 365
Ala Glu Val Tyr Ser Asp Pro Ser Lys Ile Arg Lys Glu Leu Asn Trp
370 375 380
Thr Ala Lys His Thr Asn Leu Lys Glu Ser Leu Glu Thr Ala Trp Arg
385 390 395 400
Trp Gln Lys Leu His Arg Asn Gly Tyr Gly Leu Thr Thr Ser Ser Val
405 410 415
Ser Val Tyr
<210> 224
<211> 1260
<212> DNA
<213> 拟南芥
<400> 224
atgtttagct ttggtcgtgc acgtagccag ggtcgtcaga atcgtagcat gagcttaggt 60
ggtctggatt atgcagatcc gaaaaagaaa aataactatc tgggcaaaat tctgctgacc 120
gcaagcctga ccgcactgtg catttttatg ctgaaacaga gcccgacctt taataccccg 180
agcgttttta gccgtcatga accgggtgtt acccatgttc tggttaccgg tggtgcaggt 240
tatattggta gccatgcagc actgcgtctg ctgaaagaaa gctatcgtgt taccattgtt 300
gataatctga gccgtggtaa tctggcagca gttcgtattc tgcaagaact gtttccggaa 360
ccgggtcgtc tgcagtttat ctatgccgat ctgggtgatg caaaagccgt gaataaaatc 420
tttaccgaaa atgcctttga tgccgtgatg cattttgcag cagttgcata tgttggtgaa 480
agcacccagt ttccgctgaa atattaccat aacattacca gcaataccct ggttgttctg 540
gaaaccatgg cagcacatgg tgttaaaacc ctgatttata gcagcacctg tgcaacctat 600
ggtgaaccgg atattatgcc gattaccgaa gaaacaccgc aggttccgat taatccgtat 660
ggtaaagcca aaaaaatggc cgaagatatc atcctggatt tcagcaaaaa tagcgatatg 720
gccgttatga ttctgcgcta ttttaacgtg attggtagcg atccggaagg tcgtctgggt 780
gaagcaccgc gtccggaact gcgtgaacat ggtcgtatta gcggtgcatg ttttgatgca 840
gcacgtggta ttatgcctgg tctgcagatt aaaggcaccg attacaaaac cgcagatggc 900
acctgtgttc gtgattatat tgatgttacc gatctggtgg atgcccatgt taaagcactg 960
cagaaagcaa aaccgcgtaa agtgggtatc tataatgttg gcaccggtaa aggtagcagc 1020
gttaaagaat ttgttgaggc ctgtaaaaaa gccaccggtg tggaaatcaa aatcgattat 1080
ctgcctcgtc gtgccggtga ttatgcggaa gtttatagtg atccgagcaa aattcgcaaa 1140
gaactgaatt ggaccgccaa acataccaac ctgaaagaat cactggaaac cgcatggcgt 1200
tggcagaaac tgcatcgtaa tggttatggc ctgaccacca gtagcgttag cgtttattaa 1260
<210> 225
<211> 345
<212> PRT
<213> 类志贺邻单胞菌(P. shigelloides)
<400> 225
Met Asp Ile Tyr Met Ser Arg Tyr Glu Glu Ile Thr Gln Gln Leu Ile
1 5 10 15
Phe Ser Pro Lys Thr Trp Leu Ile Thr Gly Val Ala Gly Phe Ile Gly
20 25 30
Ser Asn Leu Leu Glu Lys Leu Leu Lys Leu Asn Gln Val Val Ile Gly
35 40 45
Leu Asp Asn Phe Ser Thr Gly His Gln Tyr Asn Leu Asp Glu Val Lys
50 55 60
Thr Leu Val Ser Thr Glu Gln Trp Ser Arg Phe Cys Phe Ile Glu Gly
65 70 75 80
Asp Ile Arg Asp Leu Thr Thr Cys Glu Gln Val Met Lys Gly Val Asp
85 90 95
His Val Leu His Gln Ala Ala Leu Gly Ser Val Pro Arg Ser Ile Val
100 105 110
Asp Pro Ile Thr Thr Asn Ala Thr Asn Ile Thr Gly Phe Leu Asn Ile
115 120 125
Leu His Ala Ala Lys Asn Ala Gln Val Gln Ser Phe Thr Tyr Ala Ala
130 135 140
Ser Ser Ser Thr Tyr Gly Asp His Pro Ala Leu Pro Lys Val Glu Glu
145 150 155 160
Asn Ile Gly Asn Pro Leu Ser Pro Tyr Ala Val Thr Lys Tyr Val Asn
165 170 175
Glu Ile Tyr Ala Gln Val Tyr Ala Arg Thr Tyr Gly Phe Lys Thr Ile
180 185 190
Gly Leu Arg Tyr Phe Asn Val Phe Gly Arg Arg Gln Asp Pro Asn Gly
195 200 205
Ala Tyr Ala Ala Val Ile Pro Lys Trp Thr Ala Ala Met Leu Lys Gly
210 215 220
Asp Asp Val Tyr Ile Asn Gly Asp Gly Glu Thr Ser Arg Asp Phe Cys
225 230 235 240
Tyr Ile Asp Asn Val Ile Gln Met Asn Ile Leu Ser Ala Leu Ala Lys
245 250 255
Asp Ser Ala Lys Asp Asn Ile Tyr Asn Val Ala Val Gly Asp Arg Thr
260 265 270
Thr Leu Asn Glu Leu Ser Gly Tyr Ile Tyr Asp Glu Leu Asn Leu Ile
275 280 285
His His Ile Asp Lys Leu Ser Ile Lys Tyr Arg Glu Phe Arg Ser Gly
290 295 300
Asp Val Arg His Ser Gln Ala Asp Val Thr Lys Ala Ile Asp Leu Leu
305 310 315 320
Lys Tyr Arg Pro Asn Ile Lys Ile Arg Glu Gly Leu Arg Leu Ser Met
325 330 335
Pro Trp Tyr Val Arg Phe Leu Lys Gly
340 345
<210> 226
<211> 1038
<212> DNA
<213> 类志贺邻单胞菌
<400> 226
atggacattt atatgagccg ctatgaagaa attacccagc agctgatttt tagcccgaaa 60
acctggctga ttaccggtgt tgcaggtttt attggtagca atctgctgga aaaactgctg 120
aaactgaatc aggttgtgat tggcctggat aatttcagca ccggtcatca gtataatctg 180
gatgaagtta aaaccctggt tagcaccgaa cagtggtcac gtttttgttt tattgaaggc 240
gatattcgtg atctgaccac ctgtgaacag gttatgaaag gtgttgatca tgttctgcat 300
caggcagcac tgggtagcgt tccgcgtagc attgttgatc cgattaccac caatgcaacc 360
aatattaccg gctttctgaa tattctgcat gccgcaaaaa atgcacaggt tcagagcttt 420
acctatgcag caagcagcag cacctatggt gatcatccgg cactgccgaa agttgaagaa 480
aatattggta atccgctgag cccgtatgca gttaccaaat atgtgaatga aatttatgcc 540
caggtttacg cacgtaccta tggctttaaa accattggtc tgcgctattt caatgtgttt 600
ggtcgtcgtc aggatccgaa tggtgcatat gccgcagtta ttccgaaatg gaccgcagca 660
atgctgaaag gtgatgacgt ttatatcaat ggtgatggtg aaaccagccg tgatttttgc 720
tatattgata acgtgatcca gatgaacatt ctgagcgcac tggcaaaaga tagcgccaaa 780
gataacattt ataacgttgc agttggtgat cgtaccacac tgaatgaact gagcggttat 840
atctatgatg aactgaacct gatccaccac attgataaac tgagcatcaa atatcgcgaa 900
tttcgtagcg gtgatgttcg tcatagccag gcagatgtta ccaaagcaat tgatctgctg 960
aaatatcgtc cgaacattaa aatccgtgaa ggtctgcgtc tgagcatgcc gtggtatgtt 1020
cgttttctga aaggttaa 1038
<210> 227
<211> 520
<212> PRT
<213> 人工融合构建体
<220>
<223> 人工
<400> 227
Met Asn His Leu Arg Ala Glu Gly Pro Ala Ser Val Leu Ala Ile Gly
1 5 10 15
Thr Ala Asn Pro Glu Asn Ile Leu Leu Gln Asp Glu Phe Pro Asp Tyr
20 25 30
Tyr Phe Arg Val Thr Lys Ser Glu His Met Thr Gln Leu Lys Glu Lys
35 40 45
Phe Arg Lys Ile Cys Asp Lys Ser Met Ile Arg Lys Arg Asn Cys Phe
50 55 60
Leu Asn Glu Glu His Leu Lys Gln Asn Pro Arg Leu Val Glu His Glu
65 70 75 80
Met Gln Thr Leu Asp Ala Arg Gln Asp Met Leu Val Val Glu Val Pro
85 90 95
Lys Leu Gly Lys Asp Ala Cys Ala Lys Ala Ile Lys Glu Trp Gly Gln
100 105 110
Pro Lys Ser Lys Ile Thr His Leu Ile Phe Thr Ser Ala Ser Thr Thr
115 120 125
Asp Met Pro Gly Ala Asp Tyr His Cys Ala Lys Leu Leu Gly Leu Ser
130 135 140
Pro Ser Val Lys Arg Val Met Met Tyr Gln Leu Gly Cys Tyr Gly Gly
145 150 155 160
Gly Thr Val Leu Arg Ile Ala Lys Asp Ile Ala Glu Asn Asn Lys Gly
165 170 175
Ala Arg Val Leu Ala Val Cys Cys Asp Ile Met Ala Cys Leu Phe Arg
180 185 190
Gly Pro Ser Glu Ser Asp Leu Glu Leu Leu Val Gly Gln Ala Ile Phe
195 200 205
Gly Asp Gly Ala Ala Ala Val Ile Val Gly Ala Glu Pro Asp Glu Ser
210 215 220
Val Gly Glu Arg Pro Ile Phe Glu Leu Val Ser Thr Gly Gln Thr Ile
225 230 235 240
Leu Pro Asn Ser Glu Gly Thr Ile Gly Gly His Ile Arg Glu Ala Gly
245 250 255
Leu Ile Phe Asp Leu His Lys Asp Val Pro Met Leu Ile Ser Asn Asn
260 265 270
Ile Glu Lys Cys Leu Ile Glu Ala Phe Thr Pro Ile Gly Ile Ser Asp
275 280 285
Trp Asn Ser Ile Phe Trp Ile Thr His Pro Gly Gly Lys Ala Ile Leu
290 295 300
Asp Lys Val Glu Glu Lys Leu His Leu Lys Ser Asp Lys Phe Val Asp
305 310 315 320
Ser Arg His Val Leu Ser Glu His Gly Asn Met Ser Ser Ser Thr Val
325 330 335
Leu Phe Val Met Asp Glu Leu Arg Lys Arg Ser Leu Glu Glu Gly Lys
340 345 350
Ser Thr Thr Gly Asp Gly Phe Glu Trp Gly Val Leu Phe Gly Phe Gly
355 360 365
Pro Gly Leu Thr Val Glu Arg Val Val Val Arg Ser Val Pro Ile Lys
370 375 380
Tyr Ala Ala Thr Ser Gly Ser Thr Gly Ser Thr Gly Ser Thr Gly Ser
385 390 395 400
Gly Arg Ser Thr Gly Ser Thr Gly Ser Thr Gly Ser Gly Arg Ser His
405 410 415
Met Val Ala Val Lys His Leu Ile Val Leu Lys Phe Lys Asp Glu Ile
420 425 430
Thr Glu Ala Gln Lys Glu Glu Phe Phe Lys Thr Tyr Val Asn Leu Val
435 440 445
Asn Ile Ile Pro Ala Met Lys Asp Val Tyr Trp Gly Lys Asp Val Thr
450 455 460
Gln Lys Asn Lys Glu Glu Gly Tyr Thr His Ile Val Glu Val Thr Phe
465 470 475 480
Glu Ser Val Glu Thr Ile Gln Asp Tyr Ile Ile His Pro Ala His Val
485 490 495
Gly Phe Gly Asp Val Tyr Arg Ser Phe Trp Glu Lys Leu Leu Ile Phe
500 505 510
Asp Tyr Thr Pro Arg Lys Gly Ser
515 520
<210> 228
<211> 1563
<212> DNA
<213> 人工融合构建体
<220>
<223> 人工
<400> 228
atgaatcatt taagagctga aggtccagcc tccgttttgg ccatcggtac cgctaaccct 60
gaaaacattt tgttgcaaga cgaattccca gactactact tcagagtcac taagtccgaa 120
cacatgaccc aattgaagga gaagttcaga aagatttgtg acaagtccat gattagaaag 180
agaaactgtt tcttgaacga agaacacttg aagcaaaacc caagattggt tgaacatgaa 240
atgcaaactt tggacgctag acaagacatg ttggttgttg aagtccctaa gttgggtaag 300
gatgcctgtg ctaaggccat taaagaatgg ggtcaaccta agtccaagat tacccacttg 360
attttcacct ctgcctccac cactgacatg cctggtgctg attaccactg cgctaagtta 420
ttgggtttgt ctccatccgt taagagagtt atgatgtacc aattgggttg ctacggtggt 480
ggtactgttt taagaattgc taaggatatt gctgaaaaca acaagggtgc cagagtctta 540
gctgtctgct gtgacattat ggcttgttta ttcagaggtc catctgaatc cgacttggaa 600
ttgttggttg gtcaagctat cttcggtgac ggtgctgctg ccgttattgt tggtgctgaa 660
ccagacgaat ccgttggtga aagaccaatt tttgaattgg tttccaccgg tcaaactatt 720
ttgccaaatt ccgaaggtac catcggtggt catatcagag aagccggttt gatcttcgac 780
ttacataagg atgtcccaat gttgatctct aacaacattg aaaagtgttt gatcgaagct 840
tttaccccaa ttggtatttc tgactggaac tctatcttct ggattaccca tcctggtggt 900
aaggctattt tggataaggt cgaggaaaaa ttgcacttga agtctgacaa gttcgttgac 960
tctagacacg tcttgtccga acatggtaat atgtcctctt ccaccgtttt attcgttatg 1020
gatgagttga gaaagagatc cttagaagaa ggtaagtcca ccaccggtga tggttttgag 1080
tggggtgttt tgttcggttt cggtccaggt ttgaccgtcg aaagagttgt tgttagatct 1140
gtcccaatta agtacgcagc cacaagcggt tctacgggct ccacgggctc taccggcagt 1200
gggaggagca ctgggtcaac gggatcaaca ggtagtggaa gatcacacat ggttgccgtc 1260
aagcacttga tcgttttgaa gttcaaggat gaaatcactg aagctcaaaa ggaagaattc 1320
ttcaaaacct acgtcaactt agtcaatatt attccagcca tgaaggacgt ctattggggt 1380
aaggacgtta ctcaaaagaa taaggaggaa ggttatactc atatcgttga ggtcactttc 1440
gaatctgttg agactattca agactacatc atccacccag cccacgttgg tttcggtgat 1500
gtttatcgtt ccttctggga aaaattgttg atcttcgact acacccctag aaagggatcc 1560
taa 1563
<210> 229
<211> 381
<212> PRT
<213> A. Grandis
<400> 229
Met Ala Tyr Ser Ala Met Ala Thr Met Gly Tyr Asn Gly Met Ala Ala
1 5 10 15
Ser Cys His Thr Leu His Pro Thr Ser Pro Leu Lys Pro Phe His Gly
20 25 30
Ala Ser Thr Ser Leu Glu Ala Phe Asn Gly Glu His Met Gly Leu Leu
35 40 45
Arg Gly Tyr Ser Lys Arg Lys Leu Ser Ser Tyr Lys Asn Pro Ala Ser
50 55 60
Arg Ser Ser Asn Ala Thr Val Ala Gln Leu Leu Asn Pro Pro Gln Lys
65 70 75 80
Gly Lys Lys Ala Val Glu Phe Asp Phe Asn Lys Tyr Met Asp Ser Lys
85 90 95
Ala Met Thr Val Asn Glu Ala Leu Asn Lys Ala Ile Pro Leu Arg Tyr
100 105 110
Pro Gln Lys Ile Tyr Glu Ser Met Arg Tyr Ser Leu Leu Ala Gly Gly
115 120 125
Lys Arg Val Arg Pro Val Leu Cys Ile Ala Ala Cys Glu Leu Val Gly
130 135 140
Gly Thr Glu Glu Leu Ala Ile Pro Thr Ala Cys Ala Ile Glu Met Ile
145 150 155 160
His Thr Met Ser Leu Met His Asp Asp Leu Pro Cys Ile Asp Asn Asp
165 170 175
Asp Leu Arg Arg Gly Lys Pro Thr Asn His Lys Ile Phe Gly Glu Asp
180 185 190
Thr Ala Val Thr Ala Gly Asn Ala Leu His Ser Tyr Ala Phe Glu His
195 200 205
Ile Ala Val Ser Thr Ser Lys Thr Val Gly Ala Asp Arg Ile Leu Arg
210 215 220
Met Val Ser Glu Leu Gly Arg Ala Thr Gly Ser Glu Gly Val Met Gly
225 230 235 240
Gly Gln Met Val Asp Ile Ala Ser Glu Gly Asp Pro Ser Ile Asp Leu
245 250 255
Gln Thr Leu Glu Trp Ile His Ile His Lys Thr Ala Met Leu Leu Glu
260 265 270
Cys Ser Val Val Cys Gly Ala Ile Ile Gly Gly Ala Ser Glu Ile Val
275 280 285
Ile Glu Arg Ala Arg Arg Tyr Ala Arg Cys Val Gly Leu Leu Phe Gln
290 295 300
Val Val Asp Asp Ile Leu Asp Val Thr Lys Ser Ser Asp Glu Leu Gly
305 310 315 320
Lys Thr Ala Gly Lys Asp Leu Ile Ser Asp Lys Ala Thr Tyr Pro Lys
325 330 335
Leu Met Gly Leu Glu Lys Ala Lys Glu Phe Ser Asp Glu Leu Leu Asn
340 345 350
Arg Ala Lys Gly Glu Leu Ser Cys Phe Asp Pro Val Lys Ala Ala Pro
355 360 365
Leu Leu Gly Leu Ala Asp Tyr Val Ala Phe Arg Gln Asn
370 375 380
<210> 230
<211> 1146
<212> DNA
<213> A. Grandis
<400> 230
atggcttact ctgctatggc tactatgggt tataatggta tggctgcttc ttgtcatacc 60
ttgcatccaa cttctccatt gaaaccattt catggtgctt ccacatcttt ggaagctttt 120
aatggtgaac acatgggttt gttgagaggt tactctaaga gaaagctgtc ctcttacaaa 180
aacccagctt ctagatcttc taacgctacc gttgctcaat tattgaatcc accacaaaaa 240
ggtaagaagg ccgttgaatt tgacttcaac aagtacatgg attccaaggc tatgactgtt 300
aacgaagctt tgaacaaggc tatcccattg agatacccac aaaagatcta cgaatctatg 360
aggtactctt tgttggctgg tggtaaaagg gttagaccag ttttgtgtat tgctgcttgt 420
gaattggttg gtggtactga agaattggct attccaactg cttgtgccat tgaaatgatt 480
cacactatgt ccttgatgca cgatgatttg ccatgcattg ataacgatga cttgagaaga 540
ggtaagccaa ctaaccataa gatcttcggt gaagatactg ctgttactgc tggtaatgct 600
ttacattctt acgccttcga acatattgct gtctctactt ctaaaaccgt tggtgccgat 660
agaatcttga gaatggtttc tgaattgggt agagctactg gttctgaagg tgttatgggt 720
ggtcaaatgg ttgatattgc ttcagaaggt gatccatcca ttgacttgca aactttggaa 780
tggattcata tccataagac cgccatgttg ttggaatgtt ctgttgtttg tggtgctatt 840
attggtggtg cttctgaaat cgttattgaa agagctagaa gatacgctag atgcgttggt 900
ttgttgttcc aagttgttga tgatatcctg gatgtcacca agtcatctga tgaattaggt 960
aaaaccgctg gtaaggattt gatttctgat aaggctactt acccaaagtt gatgggttta 1020
gaaaaggcca aagaattctc cgatgagttg ttgaatagag ccaaaggtga attgtcttgt 1080
ttcgatccag ttaaggctgc tccattattg ggtttagctg attacgttgc tttcaggcaa 1140
aactaa 1146
<210> 231
<211> 541
<212> PRT
<213> 人工
<220>
<223> 人工
<400> 231
Met Ile Phe Asp Gly Thr Thr Met Ser Ile Ala Ile Gly Leu Leu Ser
1 5 10 15
Thr Leu Gly Ile Gly Ala Glu Ala Asn Pro Gln Glu Asn Phe Leu Lys
20 25 30
Cys Phe Ser Glu Tyr Ile Pro Asn Asn Pro Ala Asn Pro Lys Phe Ile
35 40 45
Tyr Thr Gln His Asp Gln Leu Tyr Met Ser Val Leu Asn Ser Thr Ile
50 55 60
Gln Asn Leu Arg Phe Thr Ser Asp Thr Thr Pro Lys Pro Leu Val Ile
65 70 75 80
Val Thr Pro Ser Asn Val Ser His Ile Gln Ala Ser Ile Leu Cys Ser
85 90 95
Lys Lys Val Gly Leu Gln Ile Arg Thr Arg Ser Gly Gly His Asp Ala
100 105 110
Glu Gly Met Ser Tyr Ile Ser Gln Val Pro Phe Val Val Val Asp Leu
115 120 125
Arg Asn Met His Ser Ile Lys Ile Asp Val His Ser Gln Thr Ala Trp
130 135 140
Val Glu Ala Gly Ala Thr Leu Gly Glu Val Tyr Tyr Trp Ile Asn Glu
145 150 155 160
Lys Asn Glu Asn Phe Ser Phe Pro Gly Gly Tyr Cys Pro Thr Val Gly
165 170 175
Val Gly Gly His Phe Ser Gly Gly Gly Tyr Gly Ala Leu Met Arg Asn
180 185 190
Tyr Gly Leu Ala Ala Asp Asn Ile Ile Asp Ala His Leu Val Asn Val
195 200 205
Asp Gly Lys Val Leu Asp Arg Lys Ser Met Gly Glu Asp Leu Phe Trp
210 215 220
Ala Ile Arg Gly Gly Gly Gly Glu Asn Phe Gly Ile Ile Ala Ala Trp
225 230 235 240
Lys Ile Lys Leu Val Ala Val Pro Ser Lys Ser Thr Ile Phe Ser Val
245 250 255
Lys Lys Asn Met Glu Ile His Gly Leu Val Lys Leu Phe Asn Lys Trp
260 265 270
Gln Asn Ile Ala Tyr Lys Tyr Asp Lys Asp Leu Val Leu Met Thr His
275 280 285
Phe Ile Thr Lys Asn Ile Thr Asp Asn His Gly Lys Asn Lys Thr Thr
290 295 300
Val His Gly Tyr Phe Ser Ser Ile Phe His Gly Gly Val Asp Ser Leu
305 310 315 320
Val Asp Leu Met Asn Lys Ser Phe Pro Glu Leu Gly Ile Lys Lys Thr
325 330 335
Asp Cys Lys Glu Phe Ser Trp Ile Asp Thr Thr Ile Phe Tyr Ser Gly
340 345 350
Val Val Asn Phe Asn Thr Ala Asn Phe Lys Lys Glu Ile Leu Leu Asp
355 360 365
Arg Ser Ala Gly Lys Lys Thr Ala Phe Ser Ile Lys Leu Asp Tyr Val
370 375 380
Lys Lys Pro Ile Pro Glu Thr Ala Met Val Lys Ile Leu Glu Lys Leu
385 390 395 400
Tyr Glu Glu Asp Val Gly Val Gly Met Tyr Val Leu Tyr Pro Tyr Gly
405 410 415
Gly Ile Met Glu Glu Ile Ser Glu Ser Ala Ile Pro Phe Pro His Arg
420 425 430
Ala Gly Ile Met Tyr Glu Leu Trp Tyr Thr Ala Ser Trp Glu Lys Gln
435 440 445
Glu Asp Asn Glu Lys His Ile Asn Trp Val Arg Ser Val Tyr Asn Phe
450 455 460
Thr Thr Pro Tyr Val Ser Gln Asn Pro Arg Leu Ala Tyr Leu Asn Tyr
465 470 475 480
Arg Asp Leu Asp Leu Gly Lys Thr Asn Pro Glu Ser Pro Asn Asn Tyr
485 490 495
Thr Gln Ala Arg Ile Trp Gly Glu Lys Tyr Phe Gly Lys Asn Phe Asn
500 505 510
Arg Leu Val Lys Val Lys Thr Lys Ala Asp Pro Asn Asn Phe Phe Arg
515 520 525
Asn Glu Gln Ser Ile Pro Pro Leu Pro Pro His His His
530 535 540
<210> 232
<211> 1626
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 232
atgattttcg atgggaccac gatgtccatt gcgatagggc tactttcaac gctgggcata 60
ggcgcagaag cgaacccgca agaaaacttt ctaaaatgct tttctgaata cattcctaac 120
aaccctgcca acccgaagtt tatctacaca caacacgatc aattgtatat gagcgtgttg 180
aatagtacaa tacagaacct gaggtttaca tccgacacaa cgccgaaacc gctagtgatc 240
gtcacaccct ccaacgtaag ccacattcag gcaagcattt tatgcagcaa gaaagtcgga 300
ctgcagataa ggacgaggtc cggaggacac gacgccgaag ggatgagcta tatctcccag 360
gtaccttttg tggtggtaga cttgagaaat atgcactcta tcaagataga cgttcactcc 420
caaaccgctt gggttgaggc gggagccacc cttggtgagg tctactactg gatcaacgaa 480
aagaatgaaa attttagctt tcctggggga tattgcccaa ctgtaggtgt tggcggccac 540
ttctcaggag gcggttatgg ggccttgatg cgtaactacg gacttgcggc cgacaacatt 600
atagacgcac atctagtgaa tgtagacggc aaagttttag acaggaagag catgggtgag 660
gatctttttt gggcaattag aggcggaggg ggagaaaatt ttggaattat cgctgcttgg 720
aaaattaagc tagttgcggt accgagcaaa agcactatat tctctgtaaa aaagaacatg 780
gagatacatg gtttggtgaa gctttttaat aagtggcaaa acatcgcgta caagtacgac 840
aaagatctgg ttctgatgac gcattttata acgaaaaata tcaccgacaa ccacggaaaa 900
aacaaaacca cagtacatgg ctacttctct agtatatttc atgggggagt cgattctctg 960
gttgatttaa tgaacaaatc attcccagag ttgggtataa agaagacaga ctgtaaggag 1020
ttctcttgga ttgacacaac tatattctat tcaggcgtag tcaactttaa cacggcgaat 1080
ttcaaaaaag agatccttct ggacagatcc gcaggtaaga aaactgcgtt ctctatcaaa 1140
ttggactatg tgaagaagcc tattcccgaa accgcgatgg tcaagatact tgagaaatta 1200
tacgaggaag atgtgggagt tggaatgtac gtactttatc cctatggtgg gataatggaa 1260
gaaatcagcg agagcgccat tccatttccc catcgtgccg gcatcatgta cgagctgtgg 1320
tatactgcga gttgggagaa gcaagaagac aacgaaaagc acattaactg ggtcagatca 1380
gtttacaatt tcaccacccc atacgtgtcc cagaatccgc gtctggctta cttgaactac 1440
cgtgatcttg acctgggtaa aacgaacccg gagtcaccca acaattacac tcaagctaga 1500
atctggggag agaaatactt tgggaagaac ttcaacaggt tagtaaaggt taaaaccaag 1560
gcagatccaa acaacttttt tagaaatgaa caatccattc ccccgctacc cccgcaccat 1620
cactaa 1626
<210> 233
<211> 540
<212> PRT
<213> 人工
<220>
<223> 人工
<400> 233
Met Ile Phe Asp Gly Thr Thr Met Ser Ile Ala Ile Gly Leu Leu Ser
1 5 10 15
Thr Leu Gly Ile Gly Ala Glu Ala Asn Pro Arg Glu Asn Phe Leu Lys
20 25 30
Cys Phe Ser Gln Tyr Ile Pro Asn Asn Ala Thr Asn Leu Lys Leu Val
35 40 45
Tyr Thr Gln Asn Asn Pro Leu Tyr Met Ser Val Leu Asn Ser Thr Ile
50 55 60
His Asn Leu Arg Phe Thr Ser Asp Thr Thr Pro Lys Pro Leu Val Ile
65 70 75 80
Val Thr Pro Ser His Val Ser His Ile Gln Gly Thr Ile Leu Cys Ser
85 90 95
Lys Lys Val Gly Leu Gln Ile Arg Thr Arg Ser Gly Gly His Asp Ser
100 105 110
Glu Gly Met Ser Tyr Ile Ser Gln Val Pro Phe Val Ile Val Asp Leu
115 120 125
Arg Asn Met Arg Ser Ile Lys Ile Asp Val His Ser Gln Thr Ala Trp
130 135 140
Val Glu Ala Gly Ala Thr Leu Gly Glu Val Tyr Tyr Trp Val Asn Glu
145 150 155 160
Lys Asn Glu Asn Leu Ser Leu Ala Ala Gly Tyr Cys Pro Thr Val Cys
165 170 175
Ala Gly Gly His Phe Gly Gly Gly Gly Tyr Gly Pro Leu Met Arg Asn
180 185 190
Tyr Gly Leu Ala Ala Asp Asn Ile Ile Asp Ala His Leu Val Asn Val
195 200 205
His Gly Lys Val Leu Asp Arg Lys Ser Met Gly Glu Asp Leu Phe Trp
210 215 220
Ala Leu Arg Gly Gly Gly Ala Glu Ser Phe Gly Ile Ile Val Ala Trp
225 230 235 240
Lys Ile Arg Leu Val Ala Val Pro Lys Ser Thr Met Phe Ser Val Lys
245 250 255
Lys Ile Met Glu Ile His Glu Leu Val Lys Leu Val Asn Lys Trp Gln
260 265 270
Asn Ile Ala Tyr Lys Tyr Asp Lys Asp Leu Leu Leu Met Thr His Phe
275 280 285
Ile Thr Arg Asn Ile Thr Asp Asn Gln Gly Lys Asn Lys Thr Ala Ile
290 295 300
His Thr Tyr Phe Ser Ser Val Phe Leu Gly Gly Val Asp Ser Leu Val
305 310 315 320
Asp Leu Met Asn Lys Ser Phe Pro Glu Leu Gly Ile Lys Lys Thr Asp
325 330 335
Cys Arg Gln Leu Ser Trp Ile Asp Thr Ile Ile Phe Tyr Ser Gly Val
340 345 350
Val Asn Tyr Asp Thr Asp Asn Phe Asn Lys Glu Ile Leu Leu Asp Arg
355 360 365
Ser Ala Gly Gln Asn Gly Ala Phe Lys Ile Lys Leu Asp Tyr Val Lys
370 375 380
Lys Pro Ile Pro Glu Ser Val Phe Val Gln Ile Leu Glu Lys Leu Tyr
385 390 395 400
Glu Glu Asp Ile Gly Ala Gly Met Tyr Ala Leu Tyr Pro Tyr Gly Gly
405 410 415
Ile Met Asp Glu Ile Ser Glu Ser Ala Ile Pro Phe Pro His Arg Ala
420 425 430
Gly Ile Leu Tyr Glu Leu Trp Tyr Ile Cys Ser Trp Glu Lys Gln Glu
435 440 445
Asp Asn Glu Lys His Leu Asn Trp Ile Arg Asn Ile Tyr Asn Phe Met
450 455 460
Thr Pro Tyr Val Ser Lys Asn Pro Arg Leu Ala Tyr Leu Asn Tyr Arg
465 470 475 480
Asp Leu Asp Ile Gly Ile Asn Asp Pro Lys Asn Pro Asn Asn Tyr Thr
485 490 495
Gln Ala Arg Ile Trp Gly Glu Lys Tyr Phe Gly Lys Asn Phe Asp Arg
500 505 510
Leu Val Lys Val Lys Thr Leu Val Asp Pro Asn Asn Phe Phe Arg Asn
515 520 525
Glu Gln Ser Ile Pro Pro Leu Pro Arg His Arg His
530 535 540
<210> 234
<211> 1623
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 234
atgatcttcg acggcacaac catgagtatc gccattggtt tgcttagcac cctgggaata 60
ggggcagaag cgaatccaag agaaaatttc ttgaagtgtt tttctcagta tatcccgaat 120
aatgcgacga accttaagtt agtatacact cagaacaacc ctctatatat gagcgttcta 180
aattctacaa tccacaacct aagatttacg tccgacacga ctccgaaacc cctagttata 240
gtgacaccgt cacatgttag ccatatacag ggcaccatac tatgttccaa aaaagttggg 300
ttacaaatac gtacccgtag cgggggacac gacagtgagg ggatgagtta tattagtcag 360
gtgcctttcg tcatagtgga tttaagaaat atgaggtcaa ttaaaatcga cgttcactca 420
caaactgcct gggttgaggc gggggccaca ttgggtgaag tatattactg ggtcaatgag 480
aagaacgaga atctttcact agcagccggt tattgtccca cagtctgcgc cggcggtcac 540
tttggcggcg gcggatacgg tcccttaatg agaaattacg ggcttgccgc agacaatatc 600
atagatgctc acttagttaa tgttcatgga aaagtgttag accgtaaaag catgggggag 660
gatctgtttt gggcgcttag agggggaggg gcagaatcat ttggaataat agtggcatgg 720
aaaatcaggc ttgtggctgt tccaaagagt accatgttct cagtaaagaa aataatggag 780
atccatgagc tagttaaact tgtgaataaa tggcaaaaca tagcctataa atatgataag 840
gacttgctgc ttatgactca tttcataacc agaaacatta cggataacca agggaagaac 900
aaaacagcca tccataccta ctttagctcc gttttcttgg gtggtgtaga cagcttagtt 960
gacctgatga acaagagttt tccggaacta ggtatcaaga agacagattg tagacaactt 1020
tcctggattg ataccataat cttttacagc ggagtcgtca attatgacac tgacaacttc 1080
aacaaggaaa ttttattaga taggagtgcg ggtcaaaatg gggccttcaa gatcaaacta 1140
gactacgtta aaaaacccat tcctgaaagt gtttttgttc agattctgga gaagctgtat 1200
gaagaagata ttggcgcggg gatgtacgct ctttatccgt acggcggcat aatggatgag 1260
attagtgaaa gcgccatccc tttcccccac agagctggta tcctgtacga gttgtggtat 1320
atctgctcct gggagaaaca ggaggataac gaaaagcact taaattggat taggaatatc 1380
tacaatttca tgacgcccta cgtttccaag aaccccaggt tggcctattt gaactacagg 1440
gatcttgata ttggaatcaa cgaccccaaa aacccaaaca actacaccca ggcaaggatt 1500
tggggagaga agtacttcgg gaagaacttc gacaggctag ttaaggtgaa aacgctagtt 1560
gatccaaata attttttcag aaacgaacag agtatccctc ccttaccgcg tcataggcac 1620
taa 1623
<210> 235
<211> 323
<212> PRT
<213> 人工
<220>
<223> 人工
<400> 235
Met Ser Ala Gly Ser Asp Gln Ile Glu Gly Ser Pro His His Glu Ser
1 5 10 15
Asp Asn Ser Ile Ala Thr Lys Ile Leu Asn Phe Gly His Thr Cys Trp
20 25 30
Lys Leu Gln Arg Pro Tyr Val Val Lys Gly Met Ile Ser Ile Ala Cys
35 40 45
Gly Leu Phe Gly Arg Glu Leu Phe Asn Asn Arg His Leu Phe Ser Trp
50 55 60
Gly Leu Met Trp Lys Ala Phe Phe Ala Leu Val Pro Ile Leu Ser Phe
65 70 75 80
Asn Phe Phe Ala Ala Ile Met Asn Gln Ile Tyr Asp Val Asp Ile Asp
85 90 95
Arg Ile Asn Lys Pro Asp Leu Pro Leu Val Ser Gly Glu Met Ser Ile
100 105 110
Glu Thr Ala Trp Ile Leu Ser Ile Ile Val Ala Leu Thr Gly Leu Ile
115 120 125
Val Thr Ile Lys Leu Lys Ser Ala Pro Leu Phe Val Phe Ile Tyr Ile
130 135 140
Phe Gly Ile Phe Ala Gly Phe Ala Tyr Ser Val Pro Pro Ile Arg Trp
145 150 155 160
Lys Gln Tyr Pro Phe Thr Asn Phe Leu Ile Thr Ile Ser Ser His Val
165 170 175
Gly Leu Ala Phe Thr Ser Tyr Ser Ala Thr Thr Ser Ala Leu Gly Leu
180 185 190
Pro Phe Val Trp Arg Pro Ala Phe Ser Phe Ile Ile Ala Phe Met Thr
195 200 205
Val Met Gly Met Thr Ile Ala Phe Ala Lys Asp Ile Ser Asp Ile Glu
210 215 220
Gly Asp Ala Lys Tyr Gly Val Ser Thr Val Ala Thr Lys Leu Gly Ala
225 230 235 240
Arg Asn Met Thr Phe Val Val Ser Gly Val Leu Leu Leu Asn Tyr Leu
245 250 255
Val Ser Ile Ser Ile Gly Ile Ile Trp Pro Gln Val Phe Lys Ser Asn
260 265 270
Ile Met Ile Leu Ser His Ala Ile Leu Ala Phe Cys Leu Ile Phe Gln
275 280 285
Thr Arg Glu Leu Ala Leu Ala Asn Tyr Ala Ser Ala Pro Ser Arg Gln
290 295 300
Phe Phe Glu Phe Ile Trp Leu Leu Tyr Tyr Ala Glu Tyr Phe Val Tyr
305 310 315 320
Val Phe Ile
<210> 236
<211> 972
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 236
atgtctgctg gctctgacca aattgaaggt tccccgcatc acgaatcaga taatagtatt 60
gccacaaaga tcttaaactt tgggcataca tgttggaaat tacaaaggcc ctacgtcgtc 120
aaaggaatga taagcatcgc ttgcggtctg ttcggaaggg aattatttaa caataggcat 180
ctattcagct gggggttaat gtggaaagct ttcttcgcgt tagtgccaat cctaagcttt 240
aactttttcg ccgccatcat gaaccagatt tatgatgttg atatcgacag gataaataag 300
ccagatcttc cattggtatc cggtgaaatg tcaatagaaa ctgcatggat attatctatt 360
atcgttgcgc tgaccggact gatagtaaca atcaaattga aatctgcacc cctgtttgtt 420
tttatatata tatttggtat tttcgctgga ttcgcttact cagtgccacc tatcaggtgg 480
aagcagtacc cattcacgaa ttttctgatc acgatctcta gccacgtcgg gttagcgttc 540
acatcttact ctgcaaccac gagtgccttg gggcttcctt tcgtctggcg tccagctttt 600
agttttatca ttgcctttat gaccgtaatg ggaatgacga tcgcattcgc aaaggacatt 660
tctgacatag agggggatgc aaaatacggt gtctccactg tggcgacaaa attaggagct 720
aggaatatga ctttcgtggt gtccggtgta ttattactaa attatctggt atctataagt 780
atcggcatca tatggccgca agtgtttaaa tccaacatta tgatactgag tcatgctatt 840
ttggcttttt gtctgatttt tcagacgcgt gagttggcgc ttgcaaacta tgcctctgcg 900
cccagcaggc agttttttga attcatatgg ttattgtact atgccgagta tttcgtctac 960
gtatttattt aa 972
<210> 237
<211> 305
<212> PRT
<213> 人工
<220>
<223> 人工
<400> 237
Met Ser Gly Ala Ala Asp Val Glu Arg Val Tyr Ala Ala Met Glu Glu
1 5 10 15
Ala Ala Gly Leu Leu Asp Val Ser Cys Ala Arg Glu Lys Ile Tyr Pro
20 25 30
Leu Leu Thr Val Phe Gln Asp Thr Leu Thr Asp Gly Val Val Val Phe
35 40 45
Ser Met Ala Ser Gly Arg Arg Ser Thr Glu Leu Asp Phe Ser Ile Ser
50 55 60
Val Pro Val Ser Gln Gly Asp Pro Tyr Ala Thr Val Val Lys Glu Gly
65 70 75 80
Leu Phe Gln Ala Thr Gly Ser Pro Val Asp Glu Leu Leu Ala Asp Thr
85 90 95
Val Ala His Leu Pro Val Ser Met Phe Ala Ile Asp Gly Glu Val Thr
100 105 110
Gly Gly Phe Lys Lys Thr Tyr Ala Phe Phe Pro Thr Asp Asp Met Pro
115 120 125
Gly Val Ala Gln Leu Ala Ala Ile Pro Ser Met Pro Ala Ser Val Ala
130 135 140
Glu Asn Ala Glu Leu Phe Ala Arg Tyr Gly Leu Asp Lys Val Gln Met
145 150 155 160
Thr Ser Met Asp Tyr Lys Lys Arg Gln Val Asn Leu Tyr Phe Ser Asp
165 170 175
Leu Lys Gln Glu Tyr Leu Gln Pro Glu Ser Val Val Ala Leu Ala Arg
180 185 190
Glu Leu Gly Leu Arg Val Pro Gly Glu Leu Gly Leu Glu Phe Cys Lys
195 200 205
Arg Ser Phe Ala Val Tyr Pro Thr Leu Asn Trp Asp Thr Gly Lys Ile
210 215 220
Asp Arg Leu Cys Phe Ala Ala Ile Ser Thr Asp Pro Thr Leu Val Pro
225 230 235 240
Ser Glu Asp Glu Arg Asp Ile Glu Met Phe Arg Asn Tyr Ala Thr Lys
245 250 255
Ala Pro Tyr Ala Tyr Val Gly Glu Lys Arg Thr Leu Val Tyr Gly Leu
260 265 270
Thr Leu Ser Ser Thr Glu Glu Tyr Tyr Lys Leu Gly Ala Tyr Tyr His
275 280 285
Ile Thr Asp Ile Gln Arg Phe Leu Leu Lys Ala Phe Asp Ala Leu Glu
290 295 300
Asp
305
<210> 238
<211> 918
<212> DNA
<213> 人工
<220>
<223> 人工
<400> 238
atgtctggtg ctgctgatgt tgaaagggtt tatgctgcta tggaagaagc tgctggtttg 60
ttggatgttt cttgtgctag agaaaagatc taccctttgt tgaccgtttt ccaagatact 120
ttgactgatg gtgttgtcgt tttctctatg gcttctggta gaagatctac tgaattggac 180
ttctccattt ccgttccagt ttctcaaggt gatccatatg ctactgttgt caaagaaggt 240
ttgtttcaag ctactggttc tccagttgat gaattattgg ctgatactgt tgctcacttg 300
ccagtttcta tgtttgctat tgatggtgaa gttaccggtg gtttcaaaaa gacttacgct 360
tttttcccaa ccgatgatat gccaggtgtt gctcaattgg ctgctattcc atctatgcca 420
gcttcagttg ctgaaaacgc tgaattattt gccagatacg gtttggataa ggtccaaatg 480
acttccatgg attacaagaa gagacaggtc aacttgtact tctccgattt gaagcaagaa 540
tacttgcaac cagaatccgt tgttgctttg gctagagaat tgggtttgag agttccaggt 600
gaattaggtt tggaattctg caagagatct ttcgctgttt acccaacttt gaattgggat 660
accggtaaga ttgatagatt gtgctttgct gctatttcca ccgatccaac tttggttcca 720
tctgaagatg aacgtgatat cgagatgttt agaaactacg ctactaaggc tccatacgct 780
tatgttggtg agaaaagaac attggtttac ggcttgactt tgtcctctac cgaagaatat 840
tacaagttgg gtgcctacta ccatatcacc gatattcaaa gattcttgct gaaggctttc 900
gatgccttgg aagattaa 918
<210> 239
<211> 722
<212> PRT
<213> 欧洲栗
<400> 239
Met Gly Lys Asn Tyr Lys Ser Leu Asp Ser Val Val Ala Ser Asp Phe
1 5 10 15
Ile Ala Leu Gly Ile Thr Ser Glu Val Ala Glu Thr Leu His Gly Arg
20 25 30
Leu Ala Glu Ile Val Cys Asn Tyr Gly Ala Ala Thr Pro Gln Thr Trp
35 40 45
Ile Asn Ile Ala Asn His Ile Leu Ser Pro Asp Leu Pro Phe Ser Leu
50 55 60
His Gln Met Leu Phe Tyr Gly Cys Tyr Lys Asp Phe Gly Pro Ala Pro
65 70 75 80
Pro Ala Trp Ile Pro Asp Pro Glu Lys Val Lys Ser Thr Asn Leu Gly
85 90 95
Ala Leu Leu Glu Lys Arg Gly Lys Glu Phe Leu Gly Val Lys Tyr Lys
100 105 110
Asp Pro Ile Ser Ser Phe Ser His Phe Gln Glu Phe Ser Val Arg Asn
115 120 125
Pro Glu Val Tyr Trp Arg Thr Val Leu Met Asp Glu Met Lys Ile Ser
130 135 140
Phe Ser Lys Asp Pro Glu Cys Ile Leu Arg Arg Asp Asp Ile Asn Asn
145 150 155 160
Pro Gly Gly Ser Glu Trp Leu Pro Gly Gly Tyr Leu Asn Ser Ala Lys
165 170 175
Asn Cys Leu Asn Val Asn Ser Asn Lys Lys Leu Asn Asp Thr Met Ile
180 185 190
Val Trp Arg Asp Glu Gly Asn Asp Asp Leu Pro Leu Asn Lys Leu Thr
195 200 205
Leu Asp Gln Leu Arg Lys Arg Val Trp Leu Val Gly Tyr Ala Leu Glu
210 215 220
Glu Met Gly Leu Glu Lys Gly Cys Ala Ile Ala Ile Asp Met Pro Met
225 230 235 240
His Val Asp Ala Val Val Ile Tyr Leu Ala Ile Val Leu Ala Gly Tyr
245 250 255
Val Val Val Ser Ile Ala Asp Ser Phe Ser Ala Pro Glu Ile Ser Thr
260 265 270
Arg Leu Arg Leu Ser Lys Ala Lys Ala Ile Phe Thr Gln Asp His Ile
275 280 285
Ile Arg Gly Lys Lys Arg Ile Pro Leu Tyr Ser Arg Val Val Glu Ala
290 295 300
Lys Ser Pro Met Ala Ile Val Ile Pro Cys Ser Gly Ser Asn Ile Gly
305 310 315 320
Ala Glu Leu Arg Asp Gly Asp Ile Ser Trp Asp Tyr Phe Leu Glu Arg
325 330 335
Ala Lys Glu Phe Lys Asn Cys Glu Phe Thr Ala Arg Glu Gln Pro Val
340 345 350
Asp Ala Tyr Thr Asn Ile Leu Phe Ser Ser Gly Thr Thr Gly Glu Pro
355 360 365
Lys Ala Ile Pro Trp Thr Gln Ala Thr Pro Leu Lys Ala Ala Ala Asp
370 375 380
Gly Trp Ser His Leu Asp Ile Arg Lys Gly Asp Val Ile Val Trp Pro
385 390 395 400
Thr Asn Leu Gly Trp Met Met Gly Pro Trp Leu Val Tyr Ala Ser Leu
405 410 415
Leu Asn Gly Ala Ser Ile Ala Leu Tyr Asn Gly Ser Pro Leu Val Ser
420 425 430
Gly Phe Ala Lys Phe Val Gln Asp Ala Lys Val Thr Met Leu Gly Val
435 440 445
Val Pro Ser Ile Val Arg Ser Trp Lys Ser Thr Asn Cys Val Ser Gly
450 455 460
Tyr Asp Trp Ser Thr Ile Arg Cys Phe Ser Ser Ser Gly Glu Ala Ser
465 470 475 480
Asn Val Asp Glu Tyr Leu Trp Leu Met Gly Arg Ala Asn Tyr Lys Pro
485 490 495
Val Ile Glu Met Cys Gly Gly Thr Glu Ile Gly Gly Ala Phe Ser Ala
500 505 510
Gly Ser Phe Leu Gln Ala Gln Ser Leu Ser Ser Phe Ser Ser Gln Cys
515 520 525
Met Gly Cys Thr Leu Tyr Ile Leu Asp Lys Asn Gly Tyr Pro Met Pro
530 535 540
Lys Asn Lys Pro Gly Ile Gly Glu Leu Ala Leu Gly Pro Val Met Phe
545 550 555 560
Gly Ala Ser Lys Thr Leu Leu Asn Gly Asn His His Asp Val Tyr Phe
565 570 575
Lys Gly Met Pro Thr Leu Asn Gly Glu Val Leu Arg Arg His Gly Asp
580 585 590
Ile Phe Glu Leu Thr Ser Asn Gly Tyr Tyr His Ala His Gly Arg Ala
595 600 605
Asp Asp Thr Met Asn Ile Gly Gly Ile Lys Ile Ser Ser Ile Glu Ile
610 615 620
Glu Arg Val Cys Asn Glu Val Asp Asp Arg Val Phe Glu Thr Thr Ala
625 630 635 640
Ile Gly Val Pro Pro Leu Gly Gly Gly Pro Glu Gln Leu Val Ile Phe
645 650 655
Phe Val Leu Lys Asp Ser Asn Asp Thr Thr Ile Asp Leu Asn Gln Leu
660 665 670
Arg Leu Ser Phe Asn Leu Gly Leu Gln Lys Lys Leu Asn Pro Leu Phe
675 680 685
Lys Val Thr Arg Val Val Pro Leu Ser Ser Leu Pro Arg Thr Ala Thr
690 695 700
Asn Lys Ile Met Arg Arg Val Leu Arg Gln Gln Phe Ser His Phe Glu
705 710 715 720
Gly Ser
<210> 240
<211> 2169
<212> DNA
<213> 欧洲栗
<400> 240
atgggtaaga attacaaatc cttggattct gttgttgctt ctgacttcat cgctttgggt 60
atcacttccg aggtcgctga aaccttacac ggtcgtttgg ctgaaattgt ttgtaactac 120
ggtgctgcta ccccacaaac ctggattaac atcgctaatc atattttgtc tccagatttg 180
ccattttctt tgcatcaaat gttgttctac ggttgttata aggatttcgg tccagctcct 240
ccagcttgga ttccagatcc agaaaaggtt aagtccacta acttgggtgc cttattggaa 300
aaaagaggta aggaattctt aggtgttaaa tacaaagacc caatctcttc tttctctcac 360
ttccaagaat tctctgttag aaacccagaa gtttactgga gaaccgtttt aatggacgag 420
atgaagatct ccttttccaa ggatccagaa tgtatcttaa gacgtgatga tattaataac 480
ccaggtggtt ccgaatggtt gccaggtggt tacttgaact ccgctaagaa ctgcttgaac 540
gttaattcca acaagaagtt aaacgacact atgatcgttt ggagggacga aggtaacgat 600
gacttgcctt tgaacaaatt aactttggac caattaagaa agagagtctg gttggttggt 660
tacgctttgg aagaaatggg tttggaaaaa ggttgtgcca ttgctatcga catgccaatg 720
cacgtcgacg ctgtcgttat ttacttggct attgtcttgg ctggttacgt tgttgtttct 780
atcgccgact ccttctccgc cccagaaatt tccactagat tgagattgtc taaggctaag 840
gccattttta cccaagatca tatcattcgt ggtaagaagc gtattccatt atactctaga 900
gtcgttgaag ctaagtctcc aatggccatt gttattccat gctctggttc caatatcggt 960
gccgaattga gggacggtga tatctcttgg gactattttt tggaaagagc taaagaattt 1020
aagaactgcg aattcaccgc cagagaacaa ccagttgacg cttacactaa catcttattc 1080
tcttctggta ccaccggtga accaaaagct attccatgga cccaagctac tcctttgaaa 1140
gccgctgctg atggttggtc ccacttagat attagaaagg gtgacgttat tgtttggcca 1200
accaacttgg gttggatgat gggtccatgg ttggtttatg cttccttgtt gaatggtgcc 1260
tccatcgctt tgtacaacgg ttctccattg gtttccggtt ttgctaagtt tgttcaagat 1320
gctaaggtca ctatgttagg tgttgttcct tctatcgtca gatcctggaa atctactaac 1380
tgtgtttctg gttacgattg gtctactatc cgttgcttct cctcttccgg tgaagcttct 1440
aacgttgacg aatatttatg gttgatgggt agagccaatt ataagcctgt cattgaaatg 1500
tgtggtggta ctgagattgg tggtgctttc tccgctggtt ccttcttgca agctcaatct 1560
ttgtcctctt tttcttctca atgtatgggt tgcactttgt acatcttgga taagaatggt 1620
tacccaatgc caaagaataa accaggtatt ggtgaattgg ccttgggtcc agttatgttc 1680
ggtgcttcca agactttatt gaacggtaac caccatgatg tttactttaa gggtatgcct 1740
actttgaacg gtgaagtttt gagaagacac ggtgacattt tcgaattaac ttccaacggt 1800
tactaccatg ctcacggtag agctgatgat accatgaaca tcggtggtat caagatctct 1860
tccattgaaa tcgagcgtgt ttgtaacgaa gttgacgaca gagttttcga aactactgcc 1920
atcggtgtcc cacctttggg tggtggtcct gaacaattgg tcattttctt cgtcttgaag 1980
gattctaacg ataccaccat cgacttgaac caattgagat tgtctttcaa cttgggtttg 2040
caaaagaagt tgaacccatt gttcaaagtc accagagttg ttccattgtc ctccttgcca 2100
cgtaccgcca ctaacaagat tatgagaaga gtcttgagac aacaattttc tcatttcgag 2160
ggatcctaa 2169
<210> 241
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 241
atctgtcaua aaacaatgtc tgactctggt ggtttcgac 39
<210> 242
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 242
cacgcgauct agtgagtgtt gttgttacac ttcc 34
<210> 243
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 243
atctgtcaua aaacaatgtc tgactctggt ggtttcgac 39
<210> 244
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 244
cacgcgauct agtgagtgtt gttgttacac ttcc 34
<210> 245
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 245
atctgtcaua aaacaatgtc tgactctggt ggtttcgac 39
<210> 246
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 246
cacgcgauct agtgagtgtt gttgttacac ttcc 34
<210> 247
<211> 41
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 247
atctgtcaua aaacaatgcc atcttctggt gacgctgctg g 41
<210> 248
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 248
cacgcgauct agttagttct acaagtacca cc 32
<210> 249
<211> 38
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 249
atctgtcaua aaacaatgat gggtgacttg actacttc 38
<210> 250
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 250
cacgcgauct atctcttcaa agaaccgatg 30
<210> 251
<211> 37
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 251
atctgtcaua aaacaatgtc ttcttctgaa ggtgttg 37
<210> 252
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 252
cacgcgauct agttagcttg agcgtttctc 30
<210> 253
<211> 37
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 253
atctgtcaua aaacaatggc tgctaacggt ggtgacc 37
<210> 254
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 254
cacgcgauct actttctttc agcgtctcta c 31
<210> 255
<211> 36
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 255
atctgtcaua aaacaatgtc tgcttctgac gctttg 36
<210> 256
<211> 34
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 256
cacgcgauct aagtctttct agaagtcttc ttcc 34
<210> 257
<211> 37
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 257
atctgtcaua aaacaatggg ttctttgact aacaacg 37
<210> 258
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 258
cacgcgauct acttagtacc agtctttcta gc 32
<210> 259
<211> 40
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 259
atctgtcaua aaacaatgga attcagattg ttgatcttgg 40
<210> 260
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 260
cacgcgauct agttcttctt caacttttca g 31
<210> 261
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 261
atctgtcaua aaacaatgac tttgttgaga gacttgttg 39
<210> 262
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 262
cacgcgauct acttagtcaa cattctgaag 30
<210> 263
<211> 38
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 263
atctgtcaua aaacaatgat cttcttctac ttcttgac 38
<210> 264
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 264
cacgcgauct agttgtcctt aaccttctta g 31
<210> 265
<211> 38
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 265
atctgtcaua aaacaatgaa cagagaagtt tctgaaag 38
<210> 266
<211> 33
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 266
cacgcgauct actttctacc gttcaattct tcc 33
<210> 267
<211> 38
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 267
atctgtcaua aaacaatgga aaagtctaac ggtttgag 38
<210> 268
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 268
cacgcgauct agaaagaaga gatgtagtcg 30
<210> 269
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 269
atctgtcaua aaacaatgtc ttctgaccca cacagaaag 39
<210> 270
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 270
cacgcgauct aagaagtgaa ttcttcgatg 30
<210> 271
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 271
atctgtcaua aaacaatgtc tacttctgaa ttggttttc 39
<210> 272
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 272
cacgcgauct agatagtaac gttagaaacg 30
<210> 273
<211> 39
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 273
atctgtcaua aaacaatgaa gcaaactgtt gttttgtac 39
<210> 274
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 274
cacgcgauct agttttgaac caagttttca ac 32
<210> 275
<211> 35
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 275
atctgtcaua aaacaatggc tagagctggt tggac 35
<210> 276
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 276
cacgcgauct agtgagtctt agacttgtga gc 32
<210> 277
<211> 38
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 277
atctgtcaua aaacaatggc ttgtactggt tggacttc 38
<210> 278
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 278
cacgcgauct agtgagtctt agacttgtga gc 32
<210> 279
<211> 35
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 279
atctgtcaua aaacaatgtc tgttaagtgg acttc 35
<210> 280
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 280
cacgcgauct agtcgttctt acccttctta g 31
<210> 281
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 281
ggatccatgt ctgactctgg tggtttcgac 30
<210> 282
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 282
aagcttctag tgagtgttgt tgttacactt cc 32
<210> 283
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 283
ggatccatgt ctgactctgg tggtttcgac 30
<210> 284
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 284
aagcttctag tgagtgttgt tgttacactt cc 32
<210> 285
<211> 30
<212> DNA
<213> 人工l
<220>
<223> 人工引物序列
<400> 285
ggatccatgt ctgactctgg tggtttcgac 30
<210> 286
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 286
aagcttctag tgagtgttgt tgttacactt cc 32
<210> 287
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 287
ggatccatgc catcttctgg tgacgctgct gg 32
<210> 288
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 288
aagcttctag ttagttctac aagtaccacc 30
<210> 289
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 289
ggatccatga tgggtgactt gactacttc 29
<210> 290
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 290
aagcttctat ctcttcaaag aaccgatg 28
<210> 291
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 291
ggatccatgt cttcttctga aggtgttg 28
<210> 292
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 292
aagcttctag ttagcttgag cgtttctc 28
<210> 293
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 293
ggatccatgg ctgctaacgg tggtgacc 28
<210> 294
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 294
aagcttctac tttctttcag cgtctctac 29
<210> 295
<211> 27
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 295
ggatccatgt ctgcttctga cgctttg 27
<210> 296
<211> 32
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 296
aagcttctaa gtctttctag aagtcttctt cc 32
<210> 297
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 297
ggatccatgg gttctttgac taacaacg 28
<210> 298
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 298
aagcttctac ttagtaccag tctttctagc 30
<210> 299
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 299
ggatccatgg aattcagatt gttgatcttg g 31
<210> 300
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 300
aagcttctag ttcttcttca acttttcag 29
<210> 301
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 301
ggatccatga ctttgttgag agacttgttg 30
<210> 302
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 302
aagcttctac ttagtcaaca ttctgaag 28
<210> 303
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 303
ggatccatga tcttcttcta cttcttgac 29
<210> 304
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 304
aagcttctag ttgtccttaa ccttcttag 29
<210> 305
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 305
ggatccatga acagagaagt ttctgaaag 29
<210> 306
<211> 31
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 306
aagcttctac tttctaccgt tcaattcttc c 31
<210> 307
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 307
ggatccatgg aaaagtctaa cggtttgag 29
<210> 308
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 308
aagcttctag aaagaagaga tgtagtcg 28
<210> 309
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 309
ggatccatgt cttctgaccc acacagaaag 30
<210> 310
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 310
aagcttctaa gaagtgaatt cttcgatg 28
<210> 311
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 311
ggatccatgt ctacttctga attggttttc 30
<210> 312
<211> 28
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 312
aagcttctag atagtaacgt tagaaacg 28
<210> 313
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 313
ggatccatga agcaaactgt tgttttgtac 30
<210> 314
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 314
aagcttctag ttttgaacca agttttcaac 30
<210> 315
<211> 26
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 315
ggatccatgg ctagagctgg ttggac 26
<210> 316
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 316
aagcttctag tgagtcttag acttgtgagc 30
<210> 317
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 317
ggatccatgg cttgtactgg ttggacttc 29
<210> 318
<211> 30
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 318
aagcttctag tgagtcttag acttgtgagc 30
<210> 319
<211> 26
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 319
ggatccatgt ctgttaagtg gacttc 26
<210> 320
<211> 29
<212> DNA
<213> 人工
<220>
<223> 人工引物序列
<400> 320
aagcttctag tcgttcttac ccttcttag 29

Claims (22)

1.一种在细胞内产生大麻素糖苷的基因修饰的微生物宿主细胞,所述细胞表达编码糖基转移酶的异源基因,所述糖基转移酶与SEQ ID NO:157或207中包括的糖基转移酶具有至少70%同一性,能够使大麻素受体与糖基供体在细胞内糖基化,从而产生所述大麻素糖苷。
2.根据权利要求2所述的基因修饰的宿主细胞,其中,所述大麻素受体是选自以下的组的大麻素苷元或大麻素糖苷:大麻色烯型(CBC)、大麻萜酚型(CBG)、大麻二酚型(CBD)、四氢大麻酚型(THC)、大麻环酚型(CBL)、大麻艾尔松型(CBE)、大麻酚型(CBN)、脱氢大麻二酚型(CBND)和二羟基大麻酚型(CBT)。
3.根据权利要求3所述的基因修饰的宿主细胞,其中,所述大麻素受体选自以下的组:大麻萜酚酸(CBGA)、大麻萜酚酸单甲基醚(CBGAM)、大麻萜酚单甲基醚(CBGM)、次大麻萜酚酸(CBGVA)、次大麻萜酚(CBGV)、大麻色烯酸(CBCA)、次大麻色烯酸(CBCVA)、次大麻色烯(CBCV)、大麻二酚酸(CBDA)、大麻二酚单甲基醚(CBDM)、大麻二酚-C4(CBD-C4)、次大麻二酚酸(CBDVA)、次大麻二酚(CBDV)、大麻二酚可尔(CBD-C1)、Δ9-反式四氢大麻酚(Δ9-THC)、Δ9-四氢大麻酚(Δ9-THC)、Δ9-顺式四氢大麻酚(Δ9-THC)、四氢大麻酚酸(THCA)、Δ9-四氢大麻酚酸A(THCA-A)、Δ9-四氢大麻酚酸B(THCA-B)、Δ9-四氢大麻酚酸-C4(THCA-C4)、Δ9-四氢大麻酚-C4(THC-C4)、Δ9-四氢次大麻酚酸(THCVA)、Δ9-四氢次大麻酚(THCV)、Δ9-四氢大麻酚可尔酸(THCA-C1)、Δ9-四氢大麻酚可尔(THC-C1)、Δ7-顺式-异-四氢次大麻酚、Δ8-四氢大麻酚酸(Δ8-THCA)、Δ8-反式-四氢大麻酚(Δ8-THC)、Δ8-四氢大麻酚(Δ8-THC)、Δ8-顺式-四氢大麻酚(Δ8-THC)、大麻环酚酸(CBLA)、大麻环酚(CBL)、次大麻环酚(CBLV)、大麻艾尔松酸A(CBEA-A)、大麻艾尔松酸B(CBEA-B)、大麻艾尔松(CBE)、cannabielsoinic acid、大麻二吡喃环烷、大麻二吡喃环烷酸、大麻酚酸(CBNA)、大麻酚甲基醚(CBNM)、大麻酚-C4(CBN-C4)、次大麻酚(CBV)、大麻酚-C2(CNB-C2)、大麻酚可尔(CBN-C1)、脱氢大麻二酚(CBND)、脱氢次大麻二酚(CBVD)、二羟基大麻酚(CBT)、10-乙氧基-9-羟基-δ-6a-四氢大麻酚、8,9-二羟基-δ-6a-四氢大麻酚、二羟基次大麻酚(CBTVE)、脱氢大麻呋喃(DCBF)、大麻呋喃(CBF)、大麻色酮(CBCN)、cannabiciuan(CBT)、10-氧亚基-δ-6a-四氢大麻酚(OTHC)、δ-9-顺式-四氢大麻酚(顺式-THC)、3,4,5,6-四氢-7-羟基-α-α-2-三甲基-9-正丙基-2,6-桥亚甲基-2H-l-苯并氧杂环辛三烯-5-甲醇(OH-异-HHCV)、大麻利比索(CBR)、三羟基-δ-9-四氢大麻酚(triOH-THC)、perrottetinene、perrottetinenic acid、11-Nor-9-羧基-THC、11-羟基-Δ9-THC、Nor-9-羧基-Δ9-四氢大麻酚、tetrahydrocannabiphorol(THCP)、cannabidiphorol(CBDP)、Cannabimovone(CBM)及其衍生物,或所述大麻素受体是选自以下组的内源性大麻素:花生四烯酰乙醇酰胺(花生四烯酰乙醇胺,AEA)、2-花生四烯酰乙醇酰胺(2-AG)、1-花生四烯酰乙醇酰胺(1-AG)和二十二碳六烯酰乙醇酰胺(DHEA,synaptamide)、油酰乙醇酰胺(OEA)、二十碳五烯酰乙醇酰胺、前列腺素乙醇酰胺、二十二碳六烯酰乙醇酰胺、亚麻酰乙醇酰胺、5(Z),8(Z),11(Z)-二十碳三烯酸乙醇酰胺(米德酸乙醇酰胺)、十七烷醇乙醇酰胺、硬脂酰乙醇酰胺、二十二碳烯酰乙醇酰胺、神经酰基乙醇酰胺、二十三酰乙醇酰胺、木蜡酰乙醇酰胺、肉豆蔻酰乙醇酰胺、十五烷酰乙醇酰胺、棕榈油酰乙醇酰胺、二十二碳六烯酸(DHA)。
4.根据前述权利要求中任一项所述的基因修饰的宿主细胞,其中所述糖基供体选自NTP-糖苷、NDP-糖苷和NMP-糖苷中的一种或多种,任选地其中核苷酸糖苷的核苷选自尿苷、腺苷、鸟苷、胞苷和脱氧胸苷,任选地其中所述糖基供体选自UDP-糖苷、ADP-糖苷、CDP-糖苷、CMP-糖苷、dTDP-糖苷和GDP-糖苷,并且任选地其中所述糖基供体选自UDP-D-葡萄糖(UDP-Glc);UDP-半乳糖(UDP-Gal);UDP-鼠李糖(UDP-Rhm)、UDP-D-木糖(UDP-Xyl);UDP-N-乙酰-D-葡糖胺(UDP-GlcNAc);UDP-N-乙酰-D-半乳糖胺(UDP-GalNAc);UDP-D-葡糖醛酸(UDP-GlcA);UDP-D-呋喃半乳糖(UDP-Galf);UDP-阿拉伯糖;UDP-芹菜糖;UDP-2-乙酰胺基-2-脱氧-α-D-甘露糖醛酸酯;UDP-N-乙酰-D-半乳糖胺4-硫酸酯;UDP-N-乙酰-D-甘露糖胺;UDP-2,3-双(3-羟基十四烷酰基)-葡糖胺;UDP-4-脱氧-4-甲酰胺基-β-L-阿拉伯吡喃糖;UDP-2,4-双(乙酰胺基)-2,4,6-三脱氧-α-D-吡喃葡萄糖;UDP-半乳糖醛酸酯;UDP-3-氨基-3-脱氧-α-D-葡萄糖;鸟苷二磷酸-D-甘露糖(GDP-Man);鸟苷二磷酸-L-岩藻糖(GDP-Fuc);鸟苷二磷酸-L-鼠李糖(GDP-Rha);胞苷单磷酸-N-乙酰神经氨酸(CMP-Neu5Ac);胞苷单磷酸-2-酮-3-脱氧-D-甘露辛酸(CMP-Kdo);和ADP-葡萄糖。
5.根据前述权利要求中任一项所述的基因修饰的宿主细胞,其中所述大麻素糖苷是选自以下的糖苷:大麻色烯型(CBC);大麻萜酚型(CBG);大麻二酚型(CBD);四氢大麻酚型(THC);大麻环酚型(CBL);大麻艾尔松型(CBE);大麻酚型(CBN);脱氢大麻二酚型(CBND)和二羟基大麻酚型(CBT),与选自以下的糖基基团连接:葡萄糖;大麻素葡糖醛酸苷;大麻素木糖苷;大麻素鼠李糖苷;大麻素半乳糖苷;大麻素N-乙酰氨基葡糖苷;大麻素N-乙酰氨基半乳糖苷和大麻素阿拉伯糖苷。
6.根据前述权利要求中任一项所述的基因修饰的宿主细胞,其中,所述大麻素糖苷选自大麻素-1'-O-β-D-葡糖苷;大麻素-1'-O-β-D-葡糖醛酸苷;大麻素-1'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖苷;大麻素-1'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖苷;大麻素-1'-O-β-D-N-乙酰半乳糖胺;大麻素-1'-O-β-D-纤维二糖苷;大麻素-1'-O-β-D-龙胆二糖苷;大麻素-1'-O-β-D-葡糖基-3'-O-β-D-葡糖苷;大麻素-1'-O-β-D-葡糖醛酸基-3'-O-β-D-葡糖醛酸苷;大麻素-1'-O-β-D-木糖基-3'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖基-3'-O-β-D-鼠李糖苷;大麻素-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;和大麻素-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺。
7.根据前述权利要求中任一项所述的基因修饰的宿主细胞,其中,所述大麻素糖苷包括通过1,4或1,6糖苷键与糖基部分共价连接的大麻素苷元或大麻素糖苷。
8.根据前述权利要求中任一项所述的基因修饰的宿主细胞,进一步包括能够产生所述大麻素受体的起作用的生物合成代谢途径,其中所述途径包含选自以下的一种或多种多肽
a)乙酰乙酰-CoA硫解酶(ACT),所述ACT将乙酰-CoA前体转化为乙酰乙酰-CoA,任选地与酿酒酵母中的天然Erg10具有至少70%同一性的ACT;
b)HMG-CoA合酶(HCS),所述HCS将乙酰乙酰-CoA前体转化为HMG-CoA,任选地与酿酒酵母中的天然Erg13具有至少70%同一性的HCS;
c)HMG-CoA还原酶(HCR),所述HCR将HMG-CoA前体转化为甲羟戊酸,任选地与酿酒酵母中的天然HMG1或HMG2具有至少70%同一性的HCR;
d)甲羟戊酸激酶(MVK),所述NVK将甲羟戊酸前体转化为甲羟戊酸-5-磷酸,任选地与酿酒酵母中的天然Erg12具有至少70%同一性的MVK;
e)磷酸甲羟戊酸激酶(PMK),所述PMK将甲羟戊酸-5-磷酸前体转化为甲羟戊酸二磷酸,任选地与酿酒酵母中的天然Erg8具有至少70%同一性的PMK;
f)甲羟戊酸焦磷酸脱羧酶(MPC),所述MPC将甲羟戊酸二磷酸前体转化为异戊烯基二磷酸(IPP),任选地与酿酒酵母中的天然MVD1具有至少70%同一性的MPC;
g)异戊烯基二磷酸/二甲基烯丙基二磷酸异构酶(IPI),所述IPI将IPP前体转化为二甲基烯丙基二磷酸(DMAPP),任选地与酿酒酵母中的天然IDI1具有至少70%同一性的IPI;
h)香叶基二磷酸合酶(GPPS),所述GPPS将IPP和DMAPP缩合成香叶基二磷酸(GPP),任选地与SEQ ID NO:45或229中包括的GPPS具有至少70%同一性的GPPS;
i)酰基活化酶(AAE),所述AAE将脂肪酸前体转化为脂肪酰基-COA,任选地与SEQ IDNO:47或239中包括的AAE具有至少70%同一性的AAE;
j)3,5,7-三氧亚基十二烷酰基-CoA合酶(TKS),所述TKS将脂肪酸-CoA前体转化为3,5,7-三氧亚基十一烷酰基-CoA,任选地与SEQ ID NO:49中包括的TKS具有至少70%同一性的TKS;
k)橄榄酸环化酶(OAC),所述OAC将3,5,7-三氧亚基十一烷酰基-CoA前体转化为divarinolic acid,任选与SEQ ID NO:51中包括的OAC具有至少70%同一性的OAC;
l)橄榄酸环化酶(OAC),所述OAC将3,5,7-三氧亚基十二烷酰基-CoA前体转化为橄榄酸,任选与SEQ ID NO:51中包括的OAC具有至少70%同一性的OAC;
m)TKS-OAC融合酶,所述TKS-OAC融合酶将脂肪酸-CoA前体转化为3,5,7-三氧亚基十一烷酰基-CoA、将3,5,7-三氧亚基十一烷酰基-CoA前体转化为divarinolic acid和将3,5,7-三氧亚基十二烷酰基-CoA前体转化为橄榄酸,任选地与SEQ ID NO 227中包括的TKS-OAC融合酶具有至少70%同一性的TKS-OAC融合酶;
n)大麻萜酚酸合酶(CBGAS),所述CBGAS将GPP和橄榄酸缩合为大麻萜酚酸(CBGA),任选地与SEQ ID NO:53、235或237中包括的CBGAS具有至少70%同一性的CBGAS;
o)大麻萜酚酸合酶(CBGAS),将GPP和divarinolic acid缩合为次大麻萜酚酸(CBGVA),任选地与SEQ ID NO:53、235或237中包括的CBGAS具有至少70%同一性的CBGAS;
p)大麻二酚酸合酶(CBDAS),所述CBDAS分别将CBGA酸和/或CBGVA转化为大麻二酚酸(CBDA)和/或次大麻二酚酸(CBDVA),任选地与SEQ ID NO:57或233中包括的CBDAS具有至少70%同一性的CBDAS;
q)四氢大麻酚酸合酶(THCAS),所述THCAS分别将CBGA和/或CBGVA转化为四氢大麻酚酸(THCA)和/或四氢次大麻酚酸(THCVA),任选地与SEQ ID NO:55或231中包括的THCAS具有至少70%同一性的THCAS;
r)大麻色烯酸合酶(CBCAS),所述CBCAS分别将CBGA和/或CBGVA转化为大麻色烯酸(CBCA)和/或次大麻色烯酸(CBCVA),任选地与SEQ ID NO:59中包括的CBCAS具有至少70%同一性的CBCAS;
s)核苷酸-葡萄糖合酶,所述核苷酸-葡萄糖合酶将蔗糖和核苷酸转化为果糖和核苷酸-葡萄糖,任选地与SEQ ID NO:209中包括的UDP-葡萄糖合酶具有至少70%同一性的UDP-葡萄糖合酶;
t)核苷酸-半乳糖4差向异构酶,所述核苷酸-半乳糖4差向异构酶将核苷酸-葡萄糖转化为核苷酸-半乳糖,任选地与SEQ ID NO:211中包括的UDP-半乳糖4差向异构酶具有至少70%同一性的UDP-半乳糖4-差向异构酶;
u)核苷酸-(葡糖醛酸)脱羧酶,所述核苷酸-(葡糖醛酸)脱羧酶将核苷酸-葡糖醛酸转化为核苷酸-木糖,任选地,与SEQ ID NO:213中包括的UDP-葡糖醛酸脱羧酶具有至少70%同一性的UDP-葡糖醛酸脱羧酶;
v)核苷酸-4-酮-6-脱氧-葡萄糖3,5差向异构酶和核苷酸-4-酮-鼠李糖4-酮-还原酶,它们一起将核苷酸-4-酮-6-脱氧-葡萄糖和NADPH转化为核苷酸-鼠李糖和NADP+,任选地与SEQ ID NO:215或219中包括的UDP-4-酮-6-脱氧-葡萄糖3,5差向异构酶具有至少70%同一性的UDP-4-酮-6-脱氧-葡萄糖3,5差向异构酶和与SEQ ID NO:215或219中包括的UDP-4-酮-鼠李糖-4-酮还原酶具有至少70%同一性的UDP-4-酮-鼠李糖-4-酮还原酶;
w)核苷酸-葡萄糖4,6脱水酶,所述核苷酸-葡萄糖4,6脱水酶将核苷酸-葡萄糖和NAD转化为核苷酸-4-酮-6-脱氧-葡萄糖和NADH,任选地与SEQ ID NO:217或219中包括的UDP-葡萄糖4,6脱水酶具有至少70%同一性的UDP-葡萄糖4,6脱水酶;
x)核苷酸-葡萄糖4,6-脱水酶和核苷酸-4-酮-6-脱氧-葡萄糖3,5差向异构酶和核苷酸-4-酮-鼠李糖-4-酮-还原酶,它们一起将核苷酸-葡萄糖和NAD+和NADPH转化为核苷酸-鼠李糖+NADH+NADP+,任选地与SEQID NO:215或219中包括的UDP-4-酮-6-脱氧-葡萄糖3,5差向异构酶具有至少70%同一性的UDP-4-酮-6-脱氧-葡萄糖3,5差向异构酶和与SEQIDNO:215或219中包括的UDP-4-酮-鼠李糖-4-酮还原酶具有至少70%同一性的UDP-4-酮-鼠李糖-4-酮还原酶以及与SEQ ID NO:217或219中包括的UDP-葡萄糖4,6脱水酶具有至少70%同一性的UDP-葡萄糖4,6脱水酶;
y)核苷酸-葡萄糖6脱氢酶,所述核苷酸-葡萄糖6脱氢酶将核苷酸-葡萄糖和2NAD+转化为核苷酸-葡糖醛酸和2NADH,任选地与SEQ ID NO:221中包括的UDP-葡萄糖6脱氢酶具有至少70%同一性的UDP-葡萄糖6脱氢酶;
z)核苷酸-阿拉伯糖4差向异构酶,所述核苷酸-阿拉伯糖4差向异构酶将核苷酸-木糖转化为核苷酸-阿拉伯糖,任选地与SEQ ID NO:223中包括的UDP-阿拉伯糖4差向异构酶具有至少70%同一性的UDP-阿拉伯糖4差向异构酶;和
aa)核苷酸-N-乙酰葡糖胺4差向异构酶,所述核苷酸-N-乙酰葡糖胺4差向异构酶将核苷酸-N-乙酰葡糖胺转化为核苷酸-N-乙酰半乳糖胺,任选地与SEQ ID NO:225中包括的UDP-N-乙酰葡糖胺4差向异构酶具有至少70%同一性的UDP-N-乙酰葡糖胺4差向异构酶。
9.一种细胞培养物,包括前述权利要求中任一项的基因修饰的宿主细胞和生长培养基。
10.一种用于产生大麻素糖苷的方法,包括在允许糖基转移酶将核苷酸糖苷的糖基部分转移到大麻素受体上的条件下,使大麻素受体同与SEQID NO:157或207中包括的糖基转移酶具有至少70%同一性的糖基转移酶和一种或多种核苷酸糖苷接触。
11.根据权利要求10所述的方法,其中糖基化在体外进行。
12.根据权利要求10所述的方法,进一步包括
a)在允许所述基因修饰的宿主细胞产生大麻素糖苷的条件下培养权利要求9所述的细胞培养物;以及
b)任选地回收和/或分离所述大麻素糖苷。
13.一种发酵液,包括权利要求9所述的细胞培养物中包括的大麻素糖苷。
14.根据权利要求13所述的发酵液,进一步包括选自以下的一种或多种化合物:
a)产生所述大麻素糖苷的起作用的生物合成代谢途径的前体或产物;
b)包括微量金属、维生素、盐、酵母氮源基础、YNB和/或氨基酸的补充营养物;并且
其中所述大麻素糖苷的浓度为至少1mg/l液体。
15.一种大麻素糖苷,包括与选自以下的糖共价连接的大麻素苷元或大麻素糖苷:木糖;鼠李糖;半乳糖;N-乙酰葡糖胺;N-乙酰半乳糖胺;和阿拉伯糖。
16.根据权利要求15所述的大麻素糖苷,其中所述大麻素糖苷选自大麻素-1'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖苷;大麻素-1'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖苷;大麻素-1'-O-β-D-N-乙酰半乳糖胺;大麻素-1'-O-β-D-纤维二糖苷;大麻素-1'-O-β-D-龙胆二糖苷;大麻素-1'-O-β-D-木糖基-3'-O-β-D-木糖苷;大麻素-1'-O-α-L-鼠李糖基-3'-O-β-D-鼠李糖苷;大麻素-1'-O-β-D-半乳糖基-3'-O-β-D-半乳糖苷;大麻素-1'-O-β-D-N-乙酰葡糖胺-3'-O-β-D-N-乙酰氨基葡糖苷;大麻素-1'-O-β-D-阿拉伯糖基-3'-O-β-D-阿拉伯糖苷;和大麻素-1'-O-β-D-N-乙酰半乳糖胺-3'-O-β-D-N-乙酰半乳糖胺。
17.一种大麻素糖苷,包括通过1,4或1,6糖苷键与糖基部分共价连接的大麻素苷元或大麻素糖苷。
18.一种组合物,包括权利要求13至14所述的发酵液和/或权利要求15至17所述的大麻素糖苷以及一种或多种剂、添加剂和/或赋形剂。
19.一种用于制备药物制剂的方法,包括将权利要求15至17所述的大麻素糖苷或权利要求18所述的组合物与一种或多种药物级赋形剂、添加剂和/或佐剂混合。
20.一种可获自权利要求19所述的方法的药物制剂。
21.一种可获自权利要求19所述的方法的药物制剂,用作药物或前药用途。
22.一种用于治疗哺乳动物中疾病的方法,包括向所述哺乳动物给药治疗有效量的权利要求20所述的药物制剂或权利要求15至17所述的大麻素糖苷。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111465700A (zh) * 2017-07-11 2020-07-28 特征生物科学公司 在酵母和植物细胞悬浮培养物中产生水溶性大麻素化合物和材料组合物

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4334441A1 (en) * 2021-05-07 2024-03-13 River Stone Biotech ApS Glycosylated opioids
WO2023044365A1 (en) * 2021-09-17 2023-03-23 Doublerainbow Biosciences Inc. Use of cyclodextrin to enhance solubility of substrates and increase enzymatic glycosylation reaction efficiency
CN114164161B (zh) * 2022-02-15 2022-05-13 佛山市汇腾生物技术有限公司 一种生产新橙皮苷的双酶共表达菌株及其构建方法和应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017053574A1 (en) * 2015-09-22 2017-03-30 Vitality Biopharma, Inc. Cannabinoid glycoside prodrugs and methods of synthesis

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PT97110B (pt) 1990-03-23 1998-11-30 Gist Brocades Nv Processo para catalisar reaccoes acelaraveis por enzimas, mediante adicao ao meio reaccional de sementes de plantas transgenicas e para obtencao das referidas sementes
US6395966B1 (en) 1990-08-09 2002-05-28 Dekalb Genetics Corp. Fertile transgenic maize plants containing a gene encoding the pat protein
AU2705895A (en) 1994-06-30 1996-01-25 Novo Nordisk Biotech, Inc. Non-toxic, non-toxigenic, non-pathogenic fusarium expression system and promoters and terminators for use therein
AU6188599A (en) 1998-10-26 2000-05-15 Novozymes A/S Constructing and screening a dna library of interest in filamentous fungal cells
CN100532561C (zh) 1999-03-22 2009-08-26 诺沃奇梅兹有限公司 新的葡糖淀粉酶及编码它的核酸序列
US7151204B2 (en) 2001-01-09 2006-12-19 Monsanto Technology Llc Maize chloroplast aldolase promoter compositions and methods for use thereof
EP2345726A3 (en) 2004-04-16 2011-12-14 DSM IP Assets B.V. Fungal promoter for expressing a gene in a fungal cell
EP1856263A1 (en) 2005-03-01 2007-11-21 DSMIP Assets B.V. Aspergillus promotors for expressing a gene in a fungal cell
WO2008098933A1 (en) 2007-02-15 2008-08-21 Dsm Ip Assets B.V. A recombinant host cell for the production of a compound of interest
EP3075848A1 (en) 2015-04-01 2016-10-05 Johann Wolfgang Goethe-Universität Frankfurt am Main Microbiological production of short fatty acids and uses thereof
BR112019019966A2 (pt) * 2017-03-24 2020-04-28 Trait Biosciences Inc biossíntese in vivo de alto nível e isolamento de canabinoides solúveis em água em sistemas de plantas
WO2019014395A1 (en) 2017-07-11 2019-01-17 Trait Biosciences, Inc. GENERATION OF WATER-SOLUBLE CANNABINOID COMPOUNDS IN PLANT CELL SUSPENSION YEAST AND CROPS AND MATERIAL COMPOSITIONS

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017053574A1 (en) * 2015-09-22 2017-03-30 Vitality Biopharma, Inc. Cannabinoid glycoside prodrugs and methods of synthesis

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
GENBANK: "PREDICTED:scopoletin glucosyltransferase-like [Cucumis sativus]", Retrieved from the Internet <URL:GENBANK ACCESSION NO. XP_011658893.1> *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111465700A (zh) * 2017-07-11 2020-07-28 特征生物科学公司 在酵母和植物细胞悬浮培养物中产生水溶性大麻素化合物和材料组合物

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