CN101522902B - 通过邻氨基苯甲酸合酶的叶绿体靶向表达产生高色氨酸玉蜀黍 - Google Patents
通过邻氨基苯甲酸合酶的叶绿体靶向表达产生高色氨酸玉蜀黍 Download PDFInfo
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Abstract
提供了编码与单体邻氨基苯甲酸合酶可操作地连接的叶绿体转运肽(CTP)的新表达载体和构建体。此外,提供了编码单体邻氨基苯甲酸合酶的新多核苷酸序列。还提供的是用于增加包含表达载体和构建体的转基因植物中的游离色氨酸水平的方法。
Description
发明背景
本申请要求于2006年8月11日提交的美国临时专利申请系列号60/837,200的优先权,所述专利申请的完整公开内容特别引入本文作为参考。
1.发明领域
本发明涉及用于表达和定位植物细胞中的邻氨基苯甲酸合酶的方法和组合物。
2.相关技术的描述
在玉蜀黍中,邻氨基苯甲酸合酶作为2亚基酶存在,其催化叶绿体中从芳族氨基酸途径到色氨酸的生物合成的第一个反应分支。它已显示是植物中的色氨酸产生调节中的重要酶。Anderson等人(美国专利号6,118,047)证实来自玉蜀黍的邻氨基苯甲酸合酶的色氨酸不敏感性α亚基的超表达导致转基因玉蜀黍植物中增加水平的色氨酸。近来,已显示来自原核来源的邻氨基苯甲酸合酶的单体形式能够增加转基因大豆和玉米中的色氨酸水平(美国专利申请系列号10/138,927,作为美国专利7,217,865授权,和10/430,011,作为美国专利申请公开20030213010公开)。
参与叶绿体内的生物合成途径的大多数蛋白质是核编码的且在细胞溶胶中合成。这些蛋白质对于质体的正确靶向因此对于其生物合成功能是必需的。在大多数情况下,这种靶向通过称为叶绿体转运肽(CTP)或质体转运肽的N末端延伸的存在来达到。来自细菌来源的转基因必须具有与编码所表达的多肽的序列融合的编码CTP序列的序列,如果所表达的多肽待区室化在叶绿体中的话。因此,外源多肽至叶绿体的转运借助于使编码CTP序列的多核苷酸序列与编码外源多肽的多核苷酸的5′区域可操作地连接来实现。
对于利用单体邻氨基苯甲酸合酶的植物细胞的操作和转化中的许多用途,将希望被引入植物细胞内的基因导致转运至质体且在质体中起作用的产物。然而,并非所有CTPs都能够以相同功效实现这种转运。用于邻氨基苯甲酸合酶在单子叶植物且特别是玉蜀黍植物中的成功表达和定位的有效和有用CTPs的鉴定是本领域需要的。
发明概述
在一个方面,本发明提供了编码包含与单体邻氨基苯甲酸合酶(AS)融合的叶绿体转运肽(CTPs)的多肽的多核苷酸,其中叶绿体转运肽能够使邻氨基苯甲酸合酶区室化在植物细胞的质体级分中。当此种邻氨基苯甲酸合酶核酸在转基因植物中表达时,升高水平的色氨酸可以在植物的细胞内达到。在本发明的一个实施方案中,提供了包含这些多核苷酸的表达载体和构建体。包含此种表达载体和构建体的重组植物细胞也是本发明的部分。通过使用本发明的序列、构建体和方法表达蛋白质获得的转基因植物细胞、种子和饲料产物进一步视为本发明的部分。
在另一个方面,本发明提供了用于增加单子叶植物中的游离色氨酸含量的方法。在一个实施方案中,该方法包括用编码包含与单体邻氨基苯甲酸合酶融合的叶绿体转运肽的多肽的多核苷酸转化单子叶植物,其中叶绿体转运肽作用于使邻氨基苯甲酸合酶活性定位或区室化在植物细胞的质体中。
在另外一个方面,本发明提供了编码来自土壤杆菌属(Agrobacterium)和中华根瘤菌属(Sinorhizobium)来源的单体邻氨基苯甲酸合酶的新型分离的多核苷酸。在本发明的一个实施方案方面中,提供了包含这些新多核苷酸的表达载体。在另外一个实施方案中,包含这些表达载体的宿主细胞、转基因植物细胞、转基因植物、来自转基因植物的种子和所得到的饲料产物也视为本发明的部分。
附图简述
图1描述了质粒pMON66560的限制性酶切图
图2描述了包含植物转化载体pMON66560的转基因玉米细胞的质体级分的蛋白质印迹分析
图3描述了质粒pMON78824的限制性酶切图
图4描述了质粒pMON78832的限制性酶切图
图5描述了质粒pMON69765的限制性酶切图
图6描述了质粒pMON69755的限制性酶切图
图7描述了质粒pMON82561的限制性酶切图
图8描述了质粒pMON78152的限制性酶切图
图9描述了质粒pMON78153的限制性酶切图
图10描述了质粒pMON82560的限制性酶切图
图11描述了质粒pMON69779的限制性酶切图
图12描述了质粒pMON78137的限制性酶切图
图13描述了质粒pMON78140的限制性酶切图
图14描述了质粒pMON78143的限制性酶切图
图15描述了质粒pMON78142的限制性酶切图
图16描述了质粒pMON78139的限制性酶切图
图17描述了质粒pMON69774的限制性酶切图
图18描述了质粒pMON69775的限制性酶切图
图19描述了质粒pMON69777的限制性酶切图
发明详述
依照本发明,提供了用于表达单体邻氨基苯甲酸合酶且将单体邻氨基苯甲酸合酶转运至植物细胞的质体的组合物。本发明特别提供了将在增加转化植物的细胞中的游离色氨酸含量中找到用途的新多核苷酸序列。此外,提供了编码来自土壤杆菌属和中华根瘤菌属的单体邻氨基苯甲酸合酶多肽的新多核苷酸。
本发明提供了编码包含与单体邻氨基苯甲酸合酶(AS)融合的叶绿体转运肽(CTPs)的多肽的多核苷酸,其中叶绿体转运肽能够使邻氨基苯甲酸合酶区室化在植物细胞的质体级分中。当此种邻氨基苯甲酸合酶核酸在转基因植物中表达时,升高水平的色氨酸可以在植物的细胞内达到。在本发明的一个方面,提供了包含这些多核苷酸的表达载体和构建体。包含此种表达载体和构建体的重组植物细胞也是本发明的部分。通过使用本发明的序列、构建体和方法表达蛋白质获得的转基因植物细胞、种子和饲料产物进一步视为本发明的部分。
在本发明的另外一个方面,提供了增加单子叶植物中的游离色氨酸含量的方法。在一个实施方案中,该方法包括用编码包含与单体邻氨基苯甲酸合酶融合的叶绿体转运肽的多肽的多核苷酸转化单子叶植物,其中叶绿体转运肽能够使邻氨基苯甲酸合酶活性区室化在植物细胞的质体中。
本发明另外涉及编码来自土壤杆菌属和中华根瘤菌属来源的单体邻氨基苯甲酸合酶的新型分离的多核苷酸。在本发明的一个方面,提供了包含这些新多核苷酸的表达载体。在另外一个实施方案中,包含这些表达载体的宿主细胞、转基因植物细胞、转基因植物、来自转基因植物的种子和所得到的饲料产物也视为本发明的部分。
显示本发明所需性状例如增加的色氨酸水平的转基因植物或种子包含赋予所需性状的插入转基因植物的基因组内的特定外源DNA。性状是与对照植物中天然存在的性状可测量的变化,所述对照植物例如具有基本上相同基因型的缺乏特定外源DNA的植物或种子。增强的所需性状可以通过使转基因植物或种子中的性状与对照植物或种子的性状相比较进行测量,所述转基因植物或种子具有与增强的所需性状相关的特定外源DNA。“高色氨酸玉蜀黍”因此指在任何植物部分优选种子中具有增加的色氨酸水平的玉米(玉蜀黍)植物;种子在本文中也可以称为仁或谷粒。
邻氨基苯甲酸合酶(AS;EC 4.1.3.27)催化植物、真菌和细菌中从芳族氨基酸途径到色氨酸的生物合成的第一个反应分支。在植物中,关于色氨酸的生物合成的化学过程被区室化在叶绿体中。因为邻氨基苯甲酸合酶是在细胞溶胶中合成的核编码的蛋白质,所以它必须通过一些方式转运到叶绿体内以参与色氨酸的生物合成。此外,对于野生型或非转基因植物天然的内源性邻氨基苯甲酸合酶通过在生物合成过程中积聚色氨酸对于反馈抑制敏感。以这种方法,非转基因植物细胞的色氨酸含量限制于相对低的水平。例如,在非转基因玉米中,色氨酸水平在植物的种子中一般小于百万分之25(ppm);通常为8-10ppm范围。本发明提供了编码与能够使邻氨基苯甲酸合酶靶向质体的叶绿体转运肽融合的来自土壤杆菌属和中华根瘤菌属的反馈不敏感的单体邻氨基苯甲酸合酶多肽的新多核苷酸。本发明进一步提供了DNA构建体和在其基因组中包含本发明的DNA构建体的至少一个植物表达盒的种子,其中种子具有比不包含构建体的种子更高的色氨酸含量。
增加的色氨酸可以通过在仁中积聚增加量(超过25ppm)的氨基酸显示于植物细胞中,且可以通过任何合适的方法进行测量,例如适当提取的组织的质谱分光光度法或高效液相层析的那种。具有增加的色氨酸的本发明的转基因玉米仁特别用作饲料或食物产物、膳食或膳食产物、或蛋白质产物、或由包含比类似变种的非转基因仁更高的色氨酸含量的仁加工的其他产物的来源。
本发明的任何植物或其部分都可以进行加工以产生饲料(例如青贮饲料)、膳食、蛋白质或油制剂。用于这种目的的特别优选的植物部分是种子。在优选实施方案中,饲料、膳食、蛋白质或油制剂被设计用于在喂养家畜(牲畜)中使用。产生饲料、膳食、蛋白质和油制剂的方法是本领域已知的,例如整体引入本文作为参考的美国专利4,957,748;5,100,679;5,219,596;5,936,069;6,005,076;6,146,669;和6,156,227。在优选实施方案中,蛋白质制剂是高蛋白制剂。高蛋白制剂优选具有超过5%w/v、更优选10%w/v和甚至更优选15%w/v的蛋白质含量。
分离的多核苷酸和多肽
在一个实施方案中,本发明提供了编码与单体邻氨基苯甲酸合酶融合的叶绿体转运肽(CTPs;质体转运肽)的分离的多核苷酸。术语“质体”意指包含前质体、白色体、造粉体、色质体和叶绿体的一类植物细胞器。在本发明的背景中,短语“转运肽”意指指导将核编码的蛋白质转运至质体的多肽。一般地,CTP或转运肽序列位于多肽的N末端处。
编码单体邻氨基苯甲酸合酶的多核苷酸和编码CTPs或质体转运肽的多核苷酸是“分离的”,因为它们已基本上与核酸在其中核酸天然存在的生物的细胞中通常与之结合的其他核酸序列分开或从其纯化出来,所述其他核酸序列即其他染色体或染色体外DNA。该术语包含生物化学纯化以便基本上去除污染核酸和其他细胞组分的核酸。该术语还包含重组核酸和化学合成的核酸。如本文所使用的,术语“基本上纯化的”指与以其天然状态通常与之结合的其他分子分开的分子。更优选地,基本上纯化的分子是在制剂中存在的占优势的种类。基本上纯化的分子可以超过60%不含、优选75%不含、更优选90%不含天然混合物中存在的其他分子(溶剂不计)。术语“基本上纯化的”不意欲包含以其天然状态存在的分子。此种分离的多核苷酸还可以是“重组的”,因为它们已与外源多核苷酸组合。例如,重组DNA分子可以是与外源启动子、或与对于宿主细胞内源的启动子可操作地连接的分离的多核苷酸。
如本文所使用的,“外源”多核苷酸是已引入宿主细胞内,且优选与其天然、未转化的状态中的细胞中存在的任何DNA序列不相同的DNA序列。“内源”或“天然”多核苷酸是天然存在于宿主细胞或生物中的DNA序列。同样地,“外源”多肽是由已引入宿主细胞内,且优选与其天然、未转化的状态中的细胞中存在的任何DNA序列不相同的分离的DNA编码的蛋白质序列。“内源”或“天然”多肽是天然存在于宿主细胞或生物中的蛋白质。
特别重要的是代表能够使单体邻氨基苯甲酸合酶多肽正确区室化在转化的植物细胞的质体中的本发明的CTP序列的多肽。CTP序列可以衍生自编码来自玉蜀黍或来自其他植物物种的质体靶向蛋白质的基因,所述其他植物物种包括但不限于芸香(Ruta graveolens)、稻(Oryzasativa)和鼠耳芥(Arabidopsis thaliana)。叶绿体转运肽序列是本领域已知的且包括下述的靶向序列:鼠耳芥核酮糖-1,5-二磷酸羧化酶(核酮糖二磷酸羧化酶-加氧酶)小亚基1(At-CTP1;Silva-Filho等人,(1996);Schnell等人,(1991);鼠耳芥5-(烯醇丙酮酰)莽草酸-3-磷酸合酶(5-(enolpyruvyl)shikimate-3-phosphate synthase)(At-CTP2;Archer等人,(1990);玉蜀黍(Zea mays)邻氨基苯甲酸合酶-α1(Zm-ASA1-CTP)和α2(Zm-ASA2-CTP)亚基;玉蜀黍二氢吡啶二羧酸合酶(Zm-DHDPS-CTP);稻ADP葡萄糖焦磷酸化酶(Os-蜡质的(Waxy)(Os-Wx)-CTP;和芸香邻氨基苯甲酸合酶α亚基(Rg-ASA短-CTP和Rg-ASA长-CTP)。关于叶绿体转运肽的用途的描述,参见都引入本文作为参考的美国专利号5,188,642和5,728,925。
编码本发明的叶绿体转运肽(CTPs)的示例性分离的多核苷酸包括包含下述核苷酸SEQ ID NOs的DNAs:
SEQ ID NO:1:编码具有修饰的切割位点(C/M)的At-CTP2(C/M)(鼠耳芥5-(烯醇丙酮酰)莽草酸-3-磷酸合酶)的核酸序列;
SEQ ID NO:2:编码具有天然的切割位点(E/K)的At-CTP2(E/K)的核酸序列;
SEQ ID NO:3:编码At-CTP2(E/K)+来自成熟的鼠耳芥属EPSPS合酶的10个氨基酸的核酸序列;
SEQ ID NO:4:编码At-CTP2(E/K)+来自成熟的鼠耳芥属EPSPS合酶的5个氨基酸的核酸序列;
SEQ ID NO:5:编码Zm-ASA1-CTP(玉蜀黍邻氨基苯甲酸合酶α1亚基)的核酸序列;
SEQ ID NO:6:编码Zm-ASA1-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α1亚基的20个氨基酸的核酸序列;
SEQ ID NO:7:编码Zm-ASA2-CTP(玉蜀黍邻氨基苯甲酸合酶α2亚基)的核酸序列;
SEQ ID NO:8:Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的5个氨基酸;
SEQ ID NO:9:编码Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的18个氨基酸的核酸序列;
SEQ ID NO:10:编码Os-Wx-CTP(稻ADP葡萄糖焦磷酸化酶)的核酸序列;
SEQ ID NO:11:编码Os-Wx-CTP+来自成熟的稻ADP葡萄糖焦磷酸化酶的5个氨基酸的核酸序列;
SEQ ID NO:12:编码Os-Wx-CTP+来自成熟的稻ADP葡萄糖焦磷酸化酶的20个氨基酸的核酸序列;
SEQ ID NO:13:编码Rg-AS短-CTP(芸香邻氨基苯甲酸合酶α亚基;Met1至Ser72)的核酸序列;
SEQ ID NO:14:编码Rg-AS长-CTP(芸香邻氨基苯甲酸合酶α亚基;Met1至Ser92)的核酸序列;
SEQ ID NO:15:编码Zm-DHDPS-CTP(玉蜀黍二氢吡啶二羧酸合酶)的核酸序列;
SEQ ID NO:16:Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的9个氨基酸;
SEQ ID NO:17:编码Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的20个氨基酸的核酸序列;
SEQ ID NO:18:编码Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的3个氨基酸的核酸序列;和
SEQ ID NO:19:编码鼠耳芥核酮糖二磷酸羧化酶-加氢酶小亚基基因叶绿体转运肽,CTP1的核酸序列。
本发明还预期编码包含例如下述氨基酸序列中的任何一个的叶绿体转运肽(CTP)的任何分离的核酸:
SEQ ID NO:20:具有修饰的切割位点(C/M)的At-CTP2(C/M)(鼠耳芥5-(烯醇丙酮酰)莽草酸-3-磷酸合酶);
SEQ ID NO:21:具有天然的切割位点(E/K)的At-CTP2(E/K);
SEQ ID NO:22 At-CTP2(E/K)+来自成熟的鼠耳芥属EPSPS合酶的10个氨基酸;
SEQ ID NO:23 At-CTP2(E/K)+来自成熟的鼠耳芥属EPSPS合酶的5个氨基酸;
SEQ ID NO:24 Zm-ASA1-CTP(玉蜀黍邻氨基苯甲酸合酶α1亚基);
SEQ ID NO:25 Zm-ASA1-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α1亚基的20个氨基酸;
SEQ ID NO:26 Zm-ASA2-CTP(玉蜀黍邻氨基苯甲酸合酶α2亚基);
SEQ ID NO:27 Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的5个氨基酸;
SEQ ID NO:28 Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的18个氨基酸;
SEQ ID NO:29 Os-Wx-CTP(稻ADP葡萄糖焦磷酸化酶);
SEQ ID NO:30 Os-Wx-CTP+来自成熟的稻ADP葡萄糖焦磷酸化酶的5个氨基酸;
SEQ ID NO:31 Os-Wx-CTP+来自成熟的稻ADP葡萄糖焦磷酸化酶的20个氨基酸;
SEQ ID NO:32 Rg-AS短-CTP(芸香邻氨基苯甲酸合酶α亚基;Met1至Ser72);
SEQ ID NO:33 Rg-AS长-CTP(芸香邻氨基苯甲酸合酶α亚基;Met1至Ser92);
SEQ ID NO:34 Zm-DHDPS-CTP(玉蜀黍二氢吡啶二羧酸合酶)
SEQ ID NO:35 Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的9个氨基酸;
SEQ ID NO:36 Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的20个氨基酸;
SEQ ID NO:37 Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的3个氨基酸;和
SEQ ID NO:38鼠耳芥核酮糖二磷酸羧化酶-加氧酶小亚基基因,CTP1。
编码与绿色荧光蛋白(GFP)、与邻氨基苯甲酸合酶、或与和GFP融合的邻氨基苯甲酸合酶融合的本发明叶绿体转运肽(CTPs)的分离的多核苷酸包括包含下述核苷酸SEQ ID NOs的DNAs:
SEQ ID NO:39编码与绿色荧光蛋白(GFP)融合的At-CTP2的核酸序列;
SEQ ID NO:40编码与绿色荧光蛋白(GFP)融合的Zm-ASA2-CTP的核酸序列;
SEQ ID NO:41编码与绿色荧光蛋白(GFP)融合的Os-Wx-CTP的核酸序列;
SEQ ID NO:42编码与绿色荧光蛋白(GFP)融合的Rg-AS短-CTP的核酸序列;
SEQ ID NO:43编码与绿色荧光蛋白(GFP)融合的Zm-DHDPS-CTP的核酸序列;
SEQ ID NO:44编码与GFP融合的Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的5个氨基酸的核酸序列;
SEQ ID NO:45编码与苜蓿根瘤菌(Rhizobium meliloti)邻氨基苯甲酸合酶融合的具有天然的切割位点E/K的At-CTP2的核酸序列;
SEQ ID NO:46编码与苜蓿根瘤菌邻氨基苯甲酸合酶融合的At-CTP2+来自成熟的鼠耳芥属EPSPS合酶的10个氨基酸的核酸序列;
SEQ ID NO:47编码与苜蓿根瘤菌邻氨基苯甲酸合酶融合的Zm-ASA2-CTP的核酸序列;
SEQ ID NO:48编码与苜蓿根瘤菌邻氨基苯甲酸合酶融合的Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的18个氨基酸的核酸序列;和
SEQ ID NO:49编码与根癌土壤杆菌(Agrobacterium tumefaciens)单体邻氨基苯甲酸合酶融合的Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基(由Zm-ASA2基因编码的前65个氨基酸)的18个氨基酸的核酸序列,所述根癌土壤杆菌单体邻氨基苯甲酸合酶与绿色荧光蛋白融合。
与绿色荧光蛋白(GFP)或邻氨基苯甲酸合酶融合的本发明的叶绿体转运肽(CTPs)的代表性序列包括包含下述多肽SEQ ID NOs的氨基酸:
SEQ ID NO:50与绿色荧光蛋白(GFP)融合的At-CTP2;
SEQ ID NO:51与绿色荧光蛋白(GFP)融合的Zm-ASA2-CTP;
SEQ ID NO:52与绿色荧光蛋白(GFP)融合的Os-Wx-CTP;
SEQ ID NO:53与绿色荧光蛋白(GFP)融合的Rg-AS短-CTP;
SEQ ID NO:54与绿色荧光蛋白(GFP)融合的Zm-DHDPS-CTP;
SEQ ID NO:55与GFP融合的Zm-DHDPS-CTP+来自成熟的玉蜀黍二氢吡啶二羧酸合酶的5个氨基酸;
SEQ ID NO:56与苜蓿根瘤菌邻氨基苯甲酸合酶融合的具有天然的切割位点E/K的At-CTP2;
SEQ ID NO:57与苜蓿根瘤菌邻氨基苯甲酸合酶融合的At-CTP2+来自成熟的鼠耳芥属EPSPS合酶的10个氨基酸;
SEQ ID NO:58与苜蓿根瘤菌邻氨基苯甲酸合酶融合的Zm-ASA2-CTP;
SEQ ID NO:59与苜蓿根瘤菌邻氨基苯甲酸合酶融合的Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的18个氨基酸;和
SEQ ID NO:60与根癌土壤杆菌单体邻氨基苯甲酸合酶融合的Zm-ASA2-CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基(由Zm-ASA2基因编码的前65个氨基酸)的18个氨基酸,所述根癌土壤杆菌单体邻氨基苯甲酸合酶与绿色荧光蛋白融合。
某些寡核苷酸对于本发明的实践也有用,例如包含SEQ ID NOs:61-198的寡核苷酸在用于瞬时原生质体测定的转染载体构建中有用;以及包含SEQ ID NOs:226-231的寡核苷酸作为PCR引物有用。
本发明的另一个方面涉及分离的单体邻氨基苯甲酸合酶(AS)及其片段;以及它们在用于获得产生升高水平的游离L-色氨酸的植物的方法中的用途。游离L-色氨酸在包含此种多肽的转基因植物中的超量产生起因于编码邻氨基苯甲酸合酶或其结构域的核酸的引入和表达。此种邻氨基苯甲酸合酶核酸包括野生型或突变α-结构域,或单体形式的邻氨基苯甲酸合酶。单体形式的邻氨基苯甲酸合酶包含在单条多肽链中的至少2个邻氨基苯甲酸合酶结构域,例如与β-结构域连接的α-结构域。
天然的植物邻氨基苯甲酸合酶一般对于经由L-色氨酸及其类似物的反馈抑制非常敏感。此种抑制构成了用于调节色氨酸合成途径的关键机制。因此,高活性、更有效或被色氨酸或其类似物抑制至较低程度的邻氨基苯甲酸合酶或其结构域将可能产生升高水平的色氨酸。根据本发明,来自根癌土壤杆菌和苜蓿中华根瘤菌(Sinorhizobium meliloti)的邻氨基苯甲酸合酶对于产生高水平的色氨酸特别有用。本发明的分离的单体邻氨基苯甲酸合酶另外包括失调(deregulated)形式及其片段。此种失调形式包括如本文所描述的中华根瘤菌属的S51C等位基因;如美国专利申请2003097677中描述的土壤杆菌属邻氨基苯甲酸合酶的F298W、V48F、V48Y、S51F和S51C等位基因;和如名称为“High TryptophanMaize”的美国专利申请11/503,532中描述的土壤杆菌属邻氨基苯甲酸合酶S51C等位基因的密码子最优化形式,所述专利申请各自整体引入本文作为参考。这些等位基因对于来自色氨酸的反馈抑制是失调的。
为了在植物或所选择的宿主细胞中产生高水平的色氨酸,分离所选择的邻氨基苯甲酸合酶核酸,且可以在体外进行操作以包括在植物细胞或其他细胞类型中基因表达所需的调节信号。因为植物中的色氨酸生物合成途径被报道存在于质体内,所以外源邻氨基苯甲酸合酶核酸被引入质体内或通过添加编码氨基末端质体转运肽的核酸区段进行修饰。此种质体转运肽可以将邻氨基苯甲酸合酶基因产物指导入质体内。
为了改变色氨酸的生物合成,编码邻氨基苯甲酸合酶活性的核酸必须被引入植物细胞或其他宿主细胞内,并且这些转化的细胞必须直接或间接进行鉴定。可以使用完整的邻氨基苯甲酸合酶或其有用部分或结构域。邻氨基苯甲酸合酶被稳定地掺入植物细胞基因组内。控制邻氨基苯甲酸合酶的表达的转录信号必须由植物细胞或其他宿主细胞识别且在植物细胞或其他宿主细胞中起作用。即,邻氨基苯甲酸合酶必须被转录成信使RNA(mRNA),并且mRNA必须在植物细胞核内是稳定的且被完整转运至细胞质用于翻译。邻氨基苯甲酸合酶mRNA必须具有由植物细胞核糖体识别且正确翻译的合适的翻译信号。多肽基因产物必须基本上逃避细胞质中的蛋白酶解攻击,被转运到正确的细胞区室内(例如质体)且能够采取将赋予酶促活性的三维构象。邻氨基苯甲酸合酶必须进一步能够在色氨酸及其衍生物的生物合成中起作用;即,它必须定位于催化生物合成中的侧面步骤(推测在质体中)的天然植物酶的附近,以获得所需底物和传递合适的产物。
即使所有这些条件都被满足,色氨酸的成功超量产生也不是可预测的事件。一些转基因的表达可能受附近的染色体元件负面影响。如果高水平的色氨酸通过突变而达到以减少反馈抑制,那么可能存在补偿邻氨基苯甲酸合酶步骤处减少的调节的其他控制机制。可能存在增加积聚的氨基酸的分解速率的机制。色氨酸和相关的氨基酸必须也在对植物无毒的水平上进行超量生产。最后,引入的性状必须是稳定的和可遗传的以允许商业开发和使用。
编码邻氨基苯甲酸合酶的多核苷酸的分离和鉴定在美国专利申请系列号:10/138,927和10/430,011中得到描述,所述美国专利申请系列号10/138,927作为美国专利申请公开20030097677公开且作为美国专利7,217,865授权;所述美国专利申请系列号10/430,011作为美国专利申请公开20030213010公开,所述专利整体引入本文作为参考。
编码本发明的邻氨基苯甲酸合酶的示例性分离的DNAs包括包含下述核苷酸SEQ ID NOs的DNAs:
SEQ ID NO:199根癌土壤杆菌野生型邻氨基苯甲酸合酶;
SEQ ID NO:201根癌土壤杆菌F298W突变等位基因;
SEQ ID NO:203根癌土壤杆菌S51F突变等位基因;
SEQ ID NO:205根癌土壤杆菌S51C突变等位基因;
SEQ ID NO:207根癌土壤杆菌密码子最优化的S51C突变等位基因;
SEQ ID NO:209苜蓿中华根瘤菌邻氨基苯甲酸合酶野生型;和
SEQ ID NO:211苜蓿中华根瘤菌邻氨基苯甲酸合酶S51C突变等位基因。
本发明还预期编码包含例如下述氨基酸序列中的任何一个的邻氨基苯甲酸合酶的分离的核酸:
SEQ ID NO:200根癌土壤杆菌野生型邻氨基苯甲酸合酶;
SEQ ID NO:202根癌土壤杆菌F298W突变体;
SEQ ID NO:204根癌土壤杆菌S51F突变体;
SEQ ID NO:206根癌土壤杆菌S51C突变体;
SEQ ID NO:208根癌土壤杆菌密码子最优化的S51C突变体;
SEQ ID NO:210苜蓿中华根瘤菌邻氨基苯甲酸合酶野生型;
SEQ ID NO:212苜蓿中华根瘤菌邻氨基苯甲酸合酶S51C突变体。
如本文就邻氨基苯甲酸合酶而言使用的,术语“单体”意指2个或更多邻氨基苯甲酸合酶结构域以功能方式并入单条多肽链内。单体邻氨基苯甲酸合酶可以在体内装配成二聚体形式。单体邻氨基苯甲酸合酶多核苷酸和多肽可以从各种生物中进行分离,例如根癌土壤杆菌、鱼腥藻属(Anabaena)M22983、巴西固氮螺菌(Azospirillum brasilense)、马尔他布鲁氏菌(Brucella melitensis)、小眼虫(Euglena gracilis)、百脉根中慢生根瘤菌(Mesorhizobium loti)、念珠蓝细菌属(Nostoc)物种PCC7120或苜蓿根瘤菌(Rhizobium meliloti)(苜蓿中华根瘤菌)。可替代地,单体邻氨基苯甲酸合酶核酸和多肽可以由选自任何方便的单体或多聚体邻氨基苯甲酸合酶基因的结构域组合进行构建。此种生物包括例如根癌土壤杆菌、鱼腥藻属M22983、鼠耳芥、巴西固氮螺菌、马尔他布鲁氏菌、百脉根中慢生根瘤菌、念珠蓝细菌属物种PCC7120、苜蓿根瘤菌(苜蓿中华根瘤菌)、血色红假单胞菌(Rhodopseudomonas palustris)、芸香、硫磺矿硫化叶菌(Sulfolobus solfataricus)、鼠伤寒沙门氏菌(Salmonella typhimurium)、粘质沙雷氏菌(Serratia marcescens)、大豆、稻、棉花、玉蜀黍、或编码邻氨基苯甲酸合酶的亚基或结构域的任何基因。编码所选择的结构域的核酸可以进行重组连接。例如,编码α-结构域的C末端的核酸可以通过形成磷酸二酯键与编码β-结构域的N末端的核酸进行连接,或反之亦然。作为替代方案,此种单结构域多肽可以进行化学连接。例如,α-结构域可以通过形成肽键经由其C末端与β-结构域的N末端进行连接,或反之亦然。
如本文所使用的,“对经由色氨酸的反馈抑制失调的”邻氨基苯甲酸合酶是,当失调的和“野生型”邻氨基苯甲酸合酶暴露于等量色氨酸或色氨酸的氨基酸类似物时,保留比相应的“野生型”或天然易感的邻氨基苯甲酸合酶多大于约10%活性的邻氨基苯甲酸合酶。优选地,失调的邻氨基苯甲酸合酶保留比相应的“野生型”或天然易感的邻氨基苯甲酸合酶多大于约20%的活性。
多肽的片段和变体也视为本发明的部分。片段是变体多肽,其具有与先前描述的多肽的氨基酸序列的部分而非全部完全相同的氨基酸序列。片段可以是“独立式的”或包含在片段形成部分或区域的较大多肽内,最优选地作为单个连续区域。优选的片段是生物学活性片段,其是介导本发明的多肽的活性的那些片段,包括具有类似活性或改善的活性或具有减少的活性的那些。还包括的是在动物中是抗原性或免疫原性的那些片段,目的是为了产生在检测法例如酶联免疫吸附测定中有用的抗体。
多肽的变体还包括通过保守氨基酸置换(残基由具有类似特征的另一个置换)与序列表中所示的序列不同的多肽。一般而言,此种置换在Ala、Val、Leu和Ile中;在Ser和Thr之间;在Asp和Glu之间;在Asn和Gln之间;在Lys和Arg之间;或在Phe和Tyr之间。特别优选的是以任何组合其中5-10;1-5;1-3或1个氨基酸是被置换、缺失或添加的变体。
显示与本文明确描述的DNA序列和氨基酸序列80%、更优选85%、甚至更优选90-95%和最优选96%-99%序列同一性的功能性邻氨基苯甲酸合酶DNA序列和功能性邻氨基苯甲酸合酶多肽也在本发明的范围内。例如,85%氨基酸同一性意指当2个序列就最大匹配进行比对时,85%的氨基酸是相同的。在使匹配达到最大中允许缺口(在匹配的2个序列的任何一个中);5或更小的缺口长度是优选的,而2或更小是更优选的。
本发明的多核苷酸可以例如用于构建用于转化植物宿主细胞的重组表达载体,如本文进一步讨论的。
植物转化载体
在本发明中重要的是多核苷酸序列或多核苷酸在重组表达载体中的使用,以指导编码与CTP融合的单体AS的多核苷酸序列在植物宿主细胞中的转录和翻译。
如本文所使用的,“重组”包括提及已通过引入异源核酸序列进行修饰的细胞或载体或衍生自如此修饰的细胞的细胞。因此,例如重组细胞表达在细胞的天然(非重组)形式内以相同形式未发现的基因,或由于有意的人为干预表达以其他方式异常表达、表达不足或根本未表达的天然基因。
术语“可操作地连接地”指启动子和第二种序列之间的功能键,其中启动子起始且介导与第二种序列对应的DNA序列的转录。如本文所使用的,“可操作地连接地”还指2个或更多不同的核苷酸序列之间的功能键,从而使得被连接的核酸序列是邻接的,且当必须连接2个蛋白质编码区时,是邻接的且在相同阅读框中。例如,使CTP编码序列与编码单体邻氨基苯甲酸合酶的核苷酸序列可操作地连接可能需要一个或多个DNA序列的操作,例如方便的限制位点或可以允许氨基末端转运序列的更好识别的接头序列。
此种多核苷酸可以根据标准PCR扩增技术使用cDNA、mRNA或基因组DNA作为模板和合适的寡核苷酸引物进行扩增。可替代地,它们可以使用标准合成技术例如自动化DNA合成仪进行合成。
表达载体最低限度包含编码在宿主细胞中表达的多肽的多核苷酸序列。一般地,表达载体置于某些调节元件的控制下,所述调节元件包括启动子、组织特异性调节元件和增强子。此种表达载体被说成与调节元件“可操作地连接”。
本发明的表达载体一般包含在植物细胞中起作用的启动子,其与编码与本发明的叶绿体转运肽(CTP)融合的邻氨基苯甲酸合酶的核酸序列和在植物宿主细胞中起作用的转录终止区可操作地连接。
本发明的示例性表达载体包括具有下述SEQ ID NOs的DNAs:
SEQ ID NO:213,编码pMON68065的核酸序列,用于Zm-ASA2-CTP+18::AgroAS(S51C)非最优化的突变等位基因的表达载体;
SEQ ID NO:214,编码pMON68066的核酸序列,用于Zm-ASA2-CTP+18::AgroAS(S51C)非天然最优化的(nno)突变等位基因的表达载体;
SEQ ID NO:215,编码pMON69757的核酸序列,用于包含AgroAS(F298W)突变等位基因的构建体的表达载体;
SEQ ID NO:216,编码pMON69770的核酸序列,用于包含具有可变3′UTR的AgroAS(S51C)非最优化的突变等位基因的构建体的表达载体;
SEQ ID NO:217,编码pMON69768的核酸序列,用于包含AgroAS(S51F)突变等位基因的构建体的表达载体;
SEQ ID NO:218,编码pMON78850的核酸序列,用于包含苜蓿根瘤菌邻氨基苯甲酸合酶野生型等位基因的构建体的表达载体;
SEQ ID NO:219,编码pMON78851的核酸序列,用于包含苜蓿根瘤菌邻氨基苯甲酸合酶S51C等位基因的构建体的表达载体。
“宿主细胞”意指包含载体且支持表达构建体的复制和/或转录或转录和翻译(表达)的细胞。用于在本发明中使用的宿主细胞可以是单子叶植物细胞或细菌细胞。本发明的细菌宿主细胞的例子是土壤杆菌属。在优选实施方案中,宿主细胞是玉蜀黍细胞。
如本文所使用的,“转基因植物”是具有稳定引入其基因组内的外源多核苷酸的植物,例如来自另一个生物的核或质体多核苷酸。
术语“种子”和“仁”理解为在含义中是等价的。术语仁经常用于描述玉米或稻植物的种子。在所有植物中,种子是由种皮、胚、糊粉和胚乳组成的成熟的胚珠。
特别重要的是本发明的多核苷酸用于制备重组表达载体以编码在宿主植物细胞中与CTP融合的单体AS的用途,其中CTP将AS的定位指导到植物宿主细胞的质体级分。植物表达构建体一般包含与本发明的核酸序列和在植物宿主细胞中起作用的转录终止区可操作地连接的在植物宿主细胞中起作用的启动子。
如本文所使用的,“启动子”意指对于从DNA的RNA转录起始所必需的DNA序列的区域。启动子位于待转录的DNA上游,且具有充当关于RNA聚合酶的结合位点的区域,且具有与其他因子一起作用以促进RNA转录的区域。更具体而言,植物中的基本启动子包含与转录起始相关的规范区域,例如CAAT和TATA框。在本发明中,优选的启动子分子和5′UTR分子允许以超过植物的其他细胞和组织的速率或水平在种子细胞或组织中转录。本领域技术人员将认识到存在在植物细胞中起作用的许多组成型和组织特异性启动子,且已在文献中得到描述。例如,启动子在下述专利中得到描述:美国专利6,437,217(玉蜀黍RS81启动子);美国专利5,641,876(稻肌动蛋白启动子);美国专利6,426,446(玉蜀黍RS324启动子);美国专利6,429,362(玉蜀黍PR-1启动子);美国专利6,232,526(玉蜀黍A3启动子);美国专利6,177,611(组成型玉蜀黍启动子);美国专利5,322,938,5,352,605、5,359,142和5,530,196(35S启动子);美国专利6,433,252(玉蜀黍L3油质蛋白启动子,P-Zm.L3);美国专利6,429,357(稻肌动蛋白2启动子以及稻肌动蛋白2内含子);美国专利5,837,848(根特异性启动子);美国专利6,294,714(光诱导型启动子);美国专利6,140,078(盐诱导型启动子);美国专利6,252,138(病原体诱导型启动子);美国专利6,175,060(磷缺乏诱导型启动子);美国专利6,635,806(γ-薏苡辛启动子,P-Cl.Gcx);美国专利申请系列号10/732,721(玉蜀黍胚特异性启动子ZmEM;emb5);美国专利7,151,204(玉蜀黍叶绿体醛缩酶启动子);SEQ ID NO:220(大麦Per1启动子);SEQID NO:221(玉蜀黍B32启动子);SEQ ID NO:222(玉蜀黍Z27启动子);SEQ ID NO:223(玉蜀黍球蛋白1启动子;Belanger和Kriz,1991);和SEQ ID NO:224(薏苡辛L-3启动子),所有这些专利都引入本文作为参考。
组成型启动子例如衍生自花椰菜花叶病毒的CaMV35S启动子(美国专利号5,858,741和5,322,938)或衍生自玄参花叶病毒的FMV35S启动子(美国专利号5,378,619)在大多数植物器官中产生高水平的表达。增强或重复形式的CaMV35S和FMV35S启动子在本发明的实践中有用,例如增强的CaMV35S(e35S)(Odell,等人,1985);美国专利5,378,619)。此外,还可以优选造成目的蛋白质在植物的特定组织中的表达,所述植物的特定组织例如叶、茎、根、块茎、种子胚乳、种子胚、果实等,并且所选择的启动子应具有所需组织和发育特异性。
调节转录终止区也可以在本发明的植物表达载体中提供。转录终止区可以由内源邻氨基苯甲酸合酶的基因序列或衍生自不同基因来源的方便的转录终止区提供。这些转录终止区通常称为3′非翻译区或3′UTRs。3′UTRs区域的例子是胭脂氨酸合酶3′区域(nos 3′;Fraley等人,1983)、小麦热激蛋白hsp17(T-Ta.Hsp17)、稻的谷蛋白基因的3′区域(Os-gt1;SEQ ID NO:225)、来自玉米醇溶蛋白基因的3′UTR例如Z273′UTR(Lopes等人,1995)、玉蜀黍球蛋白1(T-Zm.Glb1)和T-Ps.RbcS2:E9(豌豆核糖体二磷酸羧化酶-加氧酶小亚基)、WO0011200A2中公开的那些和本领域已知的其他3′UTRs,其可以与和叶绿体转运肽融合的邻氨基苯甲酸合酶编码区组合进行测试且使用。技术人员将认识到能够在单子叶植物细胞中终止转录的任何方便的转录终止区可以在本发明的构建体中使用。
此外,转录增强子或增强子的重复可以用于增加来自特定启动子的表达。此种增强子的例子包括但不限于玉蜀黍hsp70内含子(也称为Zm.DnaK)(美国专利号5,424,412Brown,等人)、Adh内含子1(Callis等人,1987)、稻肌动蛋白内含子(McElroy等人,1991;美国专利号5,641,876)、蔗糖合酶内含子(Vasil等人,1989)、TMVω元件(Gallie等人,1999)和CaMV 35S增强子或章鱼氨酸合酶增强子(美国专利号5,290,924)。因为转录起始位点和编码序列的开始之间的DNA序列即非翻译的前导序列可以影响基因表达,所以人们还可能希望采用特定前导序列。可以使用对于本领域技术人员可用的任何前导序列。优选的前导序列指导所附着基因的最佳表达水平,例如,通过增加或维持mRNA稳定性和/或通过阻止翻译的不合适的起始(Joshi,1987)。此种序列的选择任凭本领域技术人员处理。预期了衍生自在单子叶植物且特别是玉蜀黍和稻中高度表达的基因的序列。
测定本发明的分离的CTP序列使目的蛋白质靶向质体的效率的测定是本领域众所周知的。例如,报道基因例如β-葡糖醛酸糖苷酶(GUS)、氯霉素乙酰转移酶(CAT)或绿色荧光蛋白(GFP)可以与CTP序列可操作地连接。将这种基因融合物置于合适的启动子的控制下,连接到转化载体内,且转化到植物细胞内。在用于表达和定位到质体内的足够时间段后,提取质体级分且测定报道分子活性。分离的CTP序列使报道蛋白质靶向且递送至质体的能力因此得到评估且与其他已知的CTP序列进行比较;参见Silva-Filho等人,(1996)。
植物细胞转化
本发明的重组表达载体已引入其内的植物细胞、组织、器官或植物被视为转化的、转染的或转基因的。转基因的或转化的植物细胞或植物也包括植物细胞或植物的后代和由育种程序产生的后代,所述育种程序使用此种转基因植物作为杂交中的亲本且显示起因于引入的核酸序列的存在改变的表型。
在核酸序列插入细胞内的背景中的术语“引入的”意指“转染”、或“转化”或“转导”,且包括提及核酸序列整合入真核或原核细胞内,在其中核酸序列可以整合入细胞的基因组内(例如,染色体、质粒、质体或线粒体DNA),转变成自主复制子,或瞬时表达(例如,转染的mRNA)。
如本文所使用的,术语“植物”包括提及完整植物,植物器官(例如叶、茎、根等),种子和植物细胞及其后代。如本文所使用的,植物细胞包括但不限于种子悬浮培养物、胚、分生组织区域、愈伤组织、叶、根、枝条、配子体、孢子体、花粉和小孢子。可以在本发明的方法中使用的植物种类一般是单子叶植物。本发明优选的植物是玉蜀黍。
如本文所使用的,“转基因植物”包括提及在其基因组内包含异源多核苷酸的植物。一般地,异源多核苷酸稳定整合在基因组内,从而使得多核苷酸传递给连续世代。异源多核苷酸可以单独整合到基因组内或作为重组表达载体的部分整合到基因组内。“转基因的”在本文中用于包括任何细胞、细胞系、愈伤组织、组织、植物部分或植物,其基因型已通过异源核酸的存在加以改变,包括最初这样改变的那些转基因生物(transgenics)以及由起始转基因生物通过有性杂交或无性繁殖制备的那些。
因此在其细胞内具有异源多核苷酸的植物在本文中称为转基因植物。异源多核苷酸可以稳定整合到基因组内,或可以是染色体外的。优选地,本发明的多核苷酸稳定整合到基因组内从而使得多核苷酸传递给连续世代。多核苷酸单独整合到基因组内或作为重组表达载体的部分整合到基因组内。
如本文所使用的,关于核酸“异源的”是源自外来物种的核酸,或如果来自相同物种,那么通过有意的人为干预在组成和/或基因组基因座中由其天然形式相当大地修饰。例如,与异源结构基因可操作地连接的启动子来自与结构基因衍生自其的那种不同的物种,或如果来自相同物种,那么一种或两种由其最初形式相当大地修饰。异源蛋白质可以源自外来物种,或如果来自相同物种,那么通过有意的人为干预由其天然形式相当大地修饰。
用于转化宿主植物细胞的特定方法对于本发明不是关键的。植物的转化优选是持久的,即通过所引入的表达构建体整合到宿主植物基因组内,从而使得所引入的构建体传递给连续植物世代。技术人员将认识到本领域存在广泛多样的转化技术。适合于靶宿主植物的任何技术都可以在本发明的范围内使用。例如,构建体可以以多种形式引入,包括但不限于DNA链、在质粒中或在人工染色体中。构建体引入靶植物细胞内可以通过多种技术来完成,包括但不限于磷酸钙-DNA共沉淀、电穿孔、显微注射、土壤杆菌属传染、脂质体或微粒转化(即,基因枪)。
就微粒轰击(美国专利5,550,318;5,538,880;和5,610,042;所述专利各自特别整体引入本文作为参考)而言,粒子用多核苷酸包被且通过推进力递送到细胞内。示例性粒子包括包含钨、铂和优选地金的那些。用于通过粒子加速将DNA递送到植物细胞内的有用方法是Particle Delivery System(BioRad,Hercules,California),其可以用于通过筛例如不锈钢或NYTEX筛将用DNA或细胞包被的粒子推进到用在悬浮液中培养的单子叶植物细胞覆盖的过滤器表面上。微粒轰击技术可广泛应用,且可以用于转化事实上任何植物物种。可以通过微粒轰击转化的物种的例子包括单子叶植物物种,例如玉米(PCT公开WO95/06128)、大麦、小麦(整体引入本文作为参考的美国专利5,563,055)、稻、燕麦、黑麦、甘蔗和高粱;以及许多双子叶植物,包括烟草、大豆(整体引入本文作为参考的美国专利5,322,783)、向日葵、花生、棉花、番茄和一般而言的豆科植物(整体引入本文作为参考的美国专利5,563,055)。关于细节技术人员可以参考文献且选择合适的技术用于在本发明的方法中使用。
通常,包括在本发明的表达载体内的将是具有用于在宿主中表达所必需的调节区且为选择转化细胞作准备的结构基因。基因可以提供对于细胞毒剂例如抗生素、重金属、毒素等的抗性,为营养缺陷型宿主互补提供原养型,病毒免疫等。依赖于表达构建体或其组分引入其内的不同宿主物种的数目,可以使用一种或多种标记,其中用于选择的不同条件用于不同宿主。
当土壤杆菌属用于植物细胞转化时,可以使用可以引入土壤杆菌属宿主内用于与土壤杆菌属宿主中存在的T-DNA或Ti或Ri质粒同源重组的载体。包含用于重组的T-DNA的Ti或Ri质粒可以是武装的(armed)(能够引起菌瘿形成)或解除武装的(不能引起菌瘿形成),后者是允许的,只要vir基因存在于转化的土壤杆菌属宿主中。武装的质粒可以产生正常的植物细胞和菌瘿的混合物。
在一些情况下,当土壤杆菌属用作媒介物用于转化宿主植物细胞时,由T-DNA边界区为边界的表达或转录构建体将插入能够在大肠杆菌(E.coli)和土壤杆菌属中复制的广宿主范围载体内,存在在文献中描述的广宿主范围载体。常用的是pRK2或其衍生物。参见,例如,引入本文作为参考的Ditta等人,(1980)和EPA 0120515。可替代地,人们可以将待在植物细胞中表达的序列插入包含分开的复制序列的载体内,其中之一使载体在大肠杆菌中稳定,且另一个使载体在土壤杆菌属中稳定。参见,例如,McBride和Summerfelt(1990),其中利用pRiHRI(Jouanin等人,1985)复制起点且其提供植物表达载体在宿主土壤杆菌属细胞中另外的稳定性。
包括在表达构建体和T-DNA内的将是允许选择转化的土壤杆菌属和转化的宿主细胞的一种或多种标记。许多标记已开发用于与植物细胞一起使用,例如对于氯霉素、卡那霉素、氨基糖苷G418、潮霉素等的抗性。使用的特定标记对于本发明不是必需的,优选的一种或多种标记依赖于具体宿主和构建方式。
对于植物细胞使用土壤杆菌属的转化,外植体可以与转化的土壤杆菌属组合且温育足够时间用于转化,杀死细菌且在合适的选择培养基中培养植物细胞。一旦愈伤组织形成,那么枝条形成可以依照已知方法通过使用合适的植物激素进行促进,且将枝条转移至生根培养基用于植物的再生。植物随后生长至结种并且种子可以用于确立重复世代。
存在获得包含多重表达载体的本发明植物细胞的几种可能的方式。用于产生包含本发明的载体或多核苷酸序列的植物的任何方法和具有编码分开的酶的另一种多核苷酸序列的至少一种其他载体由本发明包含。例如,本发明的表达载体可以用于通过下述与第二种构建体同时转化植物,通过在单个植物转化载体(质粒)中包括2种表达载体或通过使用分开的植物转化载体,其各自表达所需基因。第二种载体可以引入已用第一种表达载体转化的植物内,或可替代地,转化的植物,一种具有第一种构建体和一种具有第二种构建体,可以使用标准育种技术进行杂交,以使构建体一起包含在相同植物中。
方法
本发明提供了增加转基因植物的种子中的游离色氨酸水平的方法。在一个实施方案中,增加色氨酸的方法包括将编码与CTP可操作地连接的单体邻氨基苯甲酸合酶的核酸序列引入植物细胞内,所述CTP能够使单体邻氨基苯甲酸合酶靶向或定位至植物细胞的叶绿体。本发明的单体邻氨基苯甲酸合酶是对经由色氨酸的反馈抑制不敏感的失调形式的酶。
如本文所使用的,植物细胞、植物组织、植物部分或植物中“增加”或“升高”水平的游离色氨酸是在未转化的植物细胞、植物组织、植物部分或植物中发现的水平的约2-200倍、优选约5-150倍、且更优选约10-100倍的水平,所述未转化的植物细胞、植物组织、植物部分或植物即其中基因组未由编码与单体邻氨基苯甲酸合酶融合的叶绿体转运肽的多核苷酸的存在改变的那种。例如,转化的植物种子中的游离L-色氨酸水平与未转化的植物种子(“原材料”)中的那些进行比较。
本发明的进一步方面是提供用于预测CTP使单体AS正确区室化至植物细胞的质体的能力的相对高通量的方法。本发明提供了利用与各种单体AS和各种CTPs融合的绿色荧光蛋白(GFP)在玉蜀黍原生质体或玉蜀黍发育中的胚中的瞬时表达的显现方法。GFP能够产生绿色荧光,在具有在395nm处的峰的UV至蓝色范围中吸收且在具有在510nm处的峰的绿色范围中发射。这种方法允许AS::GFP多肽的定位的显现。其中超过50%的定位在质体中的结果将是CTP使单体AS成功区室化的能力的阳性预示。
本发明进一步提供了制备营养上增强的玉米饲料产物的方法,其包括将本发明的玉米植物的种子加工成膳食、蛋白质或油。
此外,本发明提供了用于在转基因植物细胞中、或在衍生自此种转基因植物细胞的饲料或膳食产物中检测属于本文描述的任何CTP::AS序列组合的独特DNA序列的方法。当在DNA扩增法中测试以从由此种转基因植物细胞、或衍生自此种转基因植物细胞的饲料或膳食产物中提取的DNA扩增DNA分子时,此种转基因植物细胞、或衍生自此种转基因植物细胞的饲料或膳食产物的基因组,产生对于包含任何独特的CTP::ASDNA序列的表达载体诊断性的扩增子。如本文所使用的,“扩增子”是已使用扩增技术例如PCR或LCR合成的DNA片段。“扩增子”以其通常使用也理解为PCR产物。
本发明现在已总地进行描述,它通过参考下述实施例将更更容易理解,包括所述实施例仅用于举例说明的目的且不意欲限制本发明。
实施例
包括下述实施例以证实本发明的优选实施方案。本领域技术人员应当理解遵循由本发明人开发的代表技术的实施例中公开的技术在本发明的实践中作用良好,且因此可以被视为构成用于其实践的优选方式。然而,根据本公开内容,本领域技术人员应当理解可以在不背离本发明的概念、精神和范围的情况下,在公开的具体实施方案中进行许多改变且仍获得相同或相似结果。更具体而言,显而易见的是化学和生理学相关的某些试剂可以替代本文描述的试剂,同时将达到相同或相似结果。对于本领域技术人员显而易见的所有此种相似取代和修饰被视为在如附加权利要求限定的本发明的精神、范围和概念内。
实施例1
包含CTP1叶绿体转运肽的邻氨基苯甲酸合酶的定位
这个实施例证实编码与土壤杆菌属邻氨基苯甲酸合酶等位基因的氨基末端融合的CTP1叶绿体转运肽的转基因的蛋白质产物未有效地靶向转基因玉蜀黍中的胚质体。
植物转化载体pMON66560(图1)编码融合蛋白,其包含与AgroAS(F298W(由SEQ ID NO:201编码)的氨基末端融合的来自土壤杆菌属核酮糖二磷酸羧化酶-加氧酶小亚基基因的叶绿体转运肽序列CTP1(由SEQ ID NO:19编码),且由玉蜀黍胚特异性启动子(ZmEM)驱动。
为了从来自转基因玉蜀黍植物的未成熟的仁的胚中分离质体级分,在授粉后25-27天(DAP)时收获几个谷穗。纯合的F3转基因玉蜀黍植物包含植物转化载体pMON66560(图1)。
当其从仁中切除时,将约2.5g胚置于冰上。胚随后在冷的无菌水中漂洗3次,随后在冷PIM缓冲液(20mM Hepes/NaOH、0.5M山梨糖醇、10mM KCl、1mM MgCl2、1mM EDTA、10mM DTT,pH7.4)中漂洗。胚和随后的级分在所有分离步骤中过程保持冷。
随后将胚转移至包含5ml PIM的培养皿,且使用单边刀片精细地切碎直至切碎的胚的一致性类似沙子。使切碎的胚通过1层MiraclothTM(Calbiochem Corporation,La Jolla,CA)过滤到50ml锥形管内,且用PIM达到20ml的总体积。在分析前将这种过滤的匀浆(指定为级分H)的小等分试样贮存于-80℃。随后使过滤的匀浆在750x g下离心5分钟以使质体沉淀。倒去上清液,且将小等分试样(命名为S1)贮存于-80℃。然后向沉淀的质体中加入2.5ml PIM的等分试样,并且使用小的、软毛漆刷使沉淀重悬浮。小等分试样取出且冷冻(命名为P1的级分)后,使2.5ml重悬浮的沉淀在2个不连续的Percoll梯度中的每一个上分层。梯度管由在3ml 75%Percoll/PIM上分层的6ml 35%Percoll/PIM组成。梯度管随后在1000x g下离心8分钟。收集在35%/75%界面处所得到的质体条带且转移至另一个15ml管,并且向每个管中加入5ml 1X PIM。在750x g下离心5分钟后,取出上清液,且使2个沉淀在0.25ml PIM中重悬浮。包含纯化的质体的重悬浮的沉淀级分命名为P2且在分析前贮存于-80℃。
4种分离的级分(H、S1、P1和P2)中AS蛋白质的存在通过蛋白质印迹分析进行分析,其中使用本领域众所周知的方法。简言之,蛋白质级分通过SDS-PAGE在4-12%Bio-Rad Criterion Bis-Tris凝胶(Bio-RadLaboratories,Hercules,CA)上分离,装载18μg蛋白质/泳道。电泳后,将蛋白质转移至硝酸纤维素,且对一式两份的斑点实施用于蛋白质印迹的标准规程,包括封闭、第一抗体温育(下文描述的第一抗体)、洗涤、第二抗体温育(与辣根过氧化物酶缀合)、洗涤和化学发光检测。在印迹分析中使用的第一抗体在山羊中抗下述而激起:(a)来自玉蜀黍的邻氨基苯甲酸合酶α-2亚基,已知的质体定位蛋白质;(b)豌豆谷氨酰胺合成酶1(GS1),细胞溶胶形式的谷氨酰胺合酶(GS)(Tingey等人,1987);和(c)土壤杆菌属邻氨基苯甲酸合酶。使用本领域已知的标准方法制备分别命名为抗玉蜀黍ASα、抗豌豆GS1和抗Agro AS的抗体。
结果(图2)证实抗玉蜀黍ASα抗体尽管识别对应于所有4个级分中的玉蜀黍邻氨基苯甲酸合酶α-2亚基的条带,但在质体级分P1和P2中富集。相比之下,抗豌豆GS1抗体识别对应于级分H和S1中预期大小的蛋白质的条带,粗P1质体级分中较小量的蛋白质,和纯化的质体P2级分中几乎不能检测的量。谷氨酰胺合成酶的这种分配模式与其至细胞溶胶的定位一致。抗Agro AS抗体识别预期大小的蛋白质,其显示类似在所有4个级分中细胞溶胶标记GS1的模式,指出它也主要定位于细胞溶胶中,不管编码序列包括CTP1序列的事实。结果指出CTP1在使蛋白质区室化在玉蜀黍胚细胞的质体中的表达载体中将是没有用的。此外,结果暗示并非所有叶绿体转运肽都具有使单体AS成功定位至玉蜀黍细胞的质体的能力。
实施例2
关于邻氨基苯甲酸合酶的定位的另外转运肽序列
这个实施例描述了在原生质体转染载体的构建中整合入的各种CTP序列的设计,所述原生质体转染载体包含玉蜀黍邻氨基苯甲酸合酶-绿色荧光蛋白(Zm-AS::GFP)融合物和实施例3中详细描述的对照::GFP融合物;并且其在实施例4中描述的瞬时表达测定系统中进行评估。
表1.CTP变体
CTP名称 | SEQIDNO: | CTP的简要描述 |
At-CTP2(C/M) | 1 | 具有修饰的切割位点(C/M)的At-CTP2(鼠耳芥5-(烯醇丙酮酰)莽草酸-3-磷酸合酶) |
At-CTP2(E/K) | 2 | 具有天然的切割位点(E/K)的At-CTP2 |
CTP名称 | SEQIDNO: | CTP的简要描述 |
At-CTP2(E/K)+10 | 3 | At-CTP2+来自成熟的鼠耳介属EPSPS合酶的10个氨基酸 |
At-CTP2(E/K)+5 | 4 | At-CTP2+来自成熟的鼠耳介属EPSPS合酶的5个氨基酸 |
Zm-ASA1-CTP | 5 | Zm-ASA1 CTP(玉蜀黍邻氨基苯甲酸合酶α1亚基) |
Zm-ASA1-CTP+20 | 6 | Zm-ASA1 CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α1亚基的20个氨基酸 |
Zm-ASA2-CTP | 7 | Zm-ASA2 CTP(玉蜀黍邻氨基苯甲酸合酶α2亚基) |
Zm-ASA2-CTP+5 | 8 | Zm-ASA2 CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的5个氨基酸 |
Zm-ASA2-CTP+18 | 9 | Zm-ASA2 CTP+来自成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的18个氨基酸 |
Os-Wx-CTP | 10 | Os-Wx CTP(稻ADP葡萄糖焦磷酸化酶) |
Os-Wx-CTP+5 | 11 | Os-Wx CTP+来自成熟的稻ADP葡萄糖焦磷酸化酶的5个氨基酸 |
Os-Wx-CTP+20 | 12 | Os-Wx CTP+来自成熟的稻ADP葡萄糖焦磷酸化酶的20个氨基酸 |
Rg-AS短-CTP | 13 | Rg-AS短-CTP(芸香邻氨基苯甲酸合酶α亚基;Met1至Ser72) |
Rg-AS长-CTP | 14 | Rg-AS长-CTP(芸香邻氨基苯甲酸合酶α亚基;Met1至Ser92) |
Zm-DHDPS-CTP | 15 | Zm-DHDPS CTP(玉蜀黍二氢吡啶二羧酸合酶) |
Zm-DHDPS-CTP+9 | 16 | Zm-DHDPS来自成熟的玉蜀黍二氢吡啶二羧酸合酶的CTP+9个氨基酸 |
Zm-DHDPS-CTP+20 | 17 | Zm-DHDPS来自成熟的玉蜀黍二氢吡啶二 |
CTP名称 | SEQIDNO: | CTP的简要描述 |
羧酸合酶的CTP+20个氨基酸 | ||
Zm-DHDPS-CTP+3 | 18 | Zm-DHDPS来自成熟的玉蜀黍二氢吡啶二羧酸合酶的CTP+3个氨基酸 |
实施例3
转化载体的构建
这个实施例描述了包含玉蜀黍邻氨基苯甲酸合酶::绿色荧光蛋白(Zm-AS::GFP)融合物和对照::GFP融合物的原生质体转化载体的构建,其在实施例4中描述的瞬时原生质体和胚测定中使用。2种一般策略用于构建这些载体。
第一种策略涉及引入限制位点的CTP-AS编码序列的PCR扩增,以促进GFP编码序列的添加。第一种策略由pMON78824(图3)的构建进行例示。质粒pMON78824通过使用质粒pMON66574作为模板PCR扩增包含Zm-ASA2CTP编码序列的DNA片段进行构建。将BamHI限制位点整合入引物AS25和AS 3′(分别为SEQ ID NOs:63和61)内,以允许与在质粒pMON30098中包含的GFP编码序列符合读框地插入片段。根据TA克隆试剂盒(Invitrogen)中包含的制造商的说明书,将PCR产物克隆到pCRII载体(Invitrogen Corporation,Carlsbad,CA)内,产生质粒pMON82553。序列完整性通过DNA测序使用本领域众所周知的方法加以证实。
随后使用BamHI从pMON82553中切下Zm-ASA2 CTP片段,且插入质粒pMON30098的BamHI位点内以生成pMON78824。所得到的质粒pMON78824编码包含下述的融合蛋白(SEQ ID NO:60):a)由Zm-ASA2基因编码的前65个氨基酸;b)根癌土壤杆菌单体邻氨基苯甲酸合酶;和c)GFP编码区,全都在e35S启动子的控制下。
使用本领域众所周知的标准PCR和克隆法以相似方式构建包含各种GFP翻译融合物的另外的原生质体转化载体。转化载体IDs(pMON编号)、转化载体中包括的融合蛋白的一般描述、使用的PCR引物和PCR模板概括于表2中。
表2.构建用于原生质体转染测定的策略1质粒载体的概括
转染载体ID | 融合蛋白的一般描述 | PCR引物 | SEQIDNO: | PCR模板 |
pMON30098 | 基本载体提供用于转染载体的GFP序列 | n/a | n/a | |
pMON79960 | CTP1::GFP | CTP1 5′CTP1 3′ | 6162 | pMON66559 |
pMON79961 | CTP1::AgroAS(F298W)::GFP | CTP1 5′AS 3′ | 6163 | pMON66559 |
pMON78818 | 无CTP::AgroAS(F298W)::GFP | AS 5′AS 3′ | 6463 | pMON79961 |
pMON78820 | Zm-ASA2(包括CTP)::GFP | AS25AS23 | 6566 | pMON64201 |
pMON78822 | 无CTP:成熟的Zm-ASA2::GFP | AS2MATAS23 | 6766 | pMON64201 |
pMON78824 | Zm-ASA2-CTP+18::AgroAS(wt)::GFP | AS25AS 3′ | 6563 | pMON66574 |
pMON78140 | Os-Wx-CTP::GFP | RW-5RW-3 | 6869 | pMON66356 |
pMON78139 | Rg-AS短-CTP::GFP | Ruta-5Ruta-3 | 7071 | pMON66575 |
转染载体ID | 融合蛋白的一般描述 | PCR引物 | SEQIDNO: | PCR模板 |
pMON78143 | Rg-AS短-CTP::AgroAS(F298W)::GFP | Ruta XbaIAgroBsiWI | 7273 | pMON66575 |
pMON78142 | Rg-AS长-CTP::AgroAS(F298W)::GFP | Ruta XbaIAgroBsiWI | 7273 | pMON66571 |
pMON69763 | DHDPS+3-CTP::AgroAS(F298W)::GFP | DHDPS-67146-5′DHDPS-67146-3′ | 7475 | pMON67146 |
pMON69774 | At-CTP2(EK)::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 5′Xba1-CTP2-N13′Xba1-CTP2-N1 | 7677 | pMON78834 |
pMON69775 | At-CTP2+10::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 3′Xba1-CTP2-N23′Xba1-CTP2-N3 | 7879 | pMON78834 |
pMON69776 | Zm-ASA2-CTP::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 5′-Xba1-ZmAS2-N13′-Xba1-ZmAS2-N1 | 8081 | pMON69754 |
pMON69777 | Zm-ASA2-CTP+18::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 5′-Xba1-ZmAS1-N13′-Xba1-ZmAS2-N2 | 8283 | pMON69754 |
用于原生质体转化载体构建的第二种策略涉及对应于各种CTPs的序列的装配,和使这些序列与AgroAS(F298W)(SEQ ID NO:201)的N末端编码序列融合。CTP编码序列使用基于PCR的装配反应中的重叠引物组来产生,其基于Withers-Martinez等人(1999)的方法。这个第二种策略的例子是包含编码At-CTP2::AgroAS(F298W)::GFP融合蛋白的表达载体的质粒pMON78832(图4)的产生。
这个第二种策略的第一个步骤是合成包含AgroAS(F298W)编码序列的穿梭载体,所述编码序列进行修饰用于使各种CTPs与其N末端和GFP与其C末端符合读框地融合。为此,使用pMON79961作为模板和寡核苷酸引物AS5A(SEQ ID NO:84)和AS 3′(SEQ ID NO:63)进行PCR扩增反应,其中使用本领域众所周知的方法。所得到的包含AgroAS(F298W)编码序列的2.2kb PCR产物进行琼脂糖凝胶纯化,连接到pCRII载体内,且使用Invitrogen TA克隆试剂盒(Invitrogen)转化到感受态大肠杆菌细胞内。所得到的中间质粒命名为WDRAPP01.0005,且进行测序以证实2.2kb PCR产物的存在。随后使用BamHI从质粒WDRAPP01.0005中切下2.2kb插入片段,且克隆到质粒pBluescriptII SK+中的BamHI位点内,以产生质粒pMON82554。
这个第二种策略的第二个步骤涉及At-CTP2-AgroAS(F298W)N末端编码序列的合成。为此,制备表3中列出的下述14种寡核苷酸引物。
表3.寡核苷酸引物
寡核苷酸名称 | SEQ ID NO: |
At-CTP2-Agro N1-1 | SEQ ID NO:85 |
At-CTP2-Agro N1-2 | SEQ ID NO:86 |
At-CTP2-Agro N1-3 | SEQ ID NO:87 |
At-CTP2-Agro N1-4 | SEQ ID NO:88 |
At-CTP2-Agro N1-5 | SEQ ID NO:89 |
At-CTP2-Agro N1-6 | SEQ ID NO:90 |
At-CTP2-Agro N1-7 | SEQ ID NO:91 |
At-CTP2-Agro N1-8 | SEQ ID NO:92 |
At-CTP2-Agro N1-9 | SEQ ID NO:93 |
At-CTP2-Agro N1-10 | SEQ ID NO:94 |
At-CTP2-Agro N1-11 | SEQ ID NO:95 |
At-CTP2-Agro N1-12 | SEQ ID NO:96 |
At-CTP2-Agro N1-13 | SEQ ID NO:97 |
At-CTP2-Agro N1-14 | SEQ ID NO:98 |
通过将需要量溶解于蒸馏水中对于表5中描述的14种寡核苷酸的每一种制备母液(100μmoles/L)。随后通过使5μl每种个别的寡核苷酸溶液组合来制备寡核苷酸混合物(At-CTP2 N1寡核苷酸混合物)。使用下述条件进行PCR扩增。
第一次PCR
混合物1:
1微升CTP2N1寡核苷酸混合物
1微升4dNTP混合物(Roche,各10毫摩尔)
23微升水
混合物2:
6微升25mM MgCl2
5微升PCR缓冲液(不含MgCl2)
0.75微升Expand Hi-Fi酶混合物(Roche)
13.25微升水
使混合物1和混合物2组合在薄壁PCR管中且如下进行PCR反应:
1)94℃/2分钟
2)94℃/30秒
3)45℃/30秒
4)72℃/30秒
5)回到步骤2再4次
6)72℃/2分钟
7)4℃/保持
第二次PCR
混合物3:
1微升第一次PCR反应物
1.5微升N1-1寡核苷酸(10皮摩尔/微升)
1.5微升N1-14寡核苷酸(10皮摩尔/微升)
1微升dNTP混合物
20微升水
混合物4:
6微升25mM MgCl2
5微升PCR缓冲液(不含MgCl2)
0.75微升Expand Hi-Fi酶混合物(Roche)
13.25微升水
使混合物1和混合物2组合在薄壁PCR管中且如下进行PCR反应:
使混合物3和混合物4组合在薄壁PCR管中且如下进行PCR反应:
1)94℃/2分钟
2)94℃/30秒
3)55℃/30秒
4)72℃/30秒
5)回到步骤2再24次
6)72℃/2分钟
7)4℃/保持
如上所述所得到的正确大小(~0.3kb)的PCR产物进行琼脂糖凝胶纯化,且连接到pCRII载体(Invitrogen TA克隆试剂盒,Invitrogen)内。证实序列后,用NcoI和BsiWI消化质粒且克隆到pMON82554内,替代先前已在NcoI和BsiWI位点处取出的片段。所得到的中间质粒随后用BamH1进行消化,以产生随后克隆到pMON30098内的片段。这种所得到的质粒载体包含e35S启动子::hsp70内含子::At-CTP2:AgroAS(F298W)::GFP::nos 3’UTR遗传元件(pMON78832;图4)。
每种CTP的几个变体以上文描述的方式通过在CTP的3′末端处添加编码3-20个氨基酸的相应成熟蛋白质的DNA序列进行构建。对于包含CTP变体的每种pCRII载体,随后如上所述从载体中取出pCRII载体的NcoI和BSiWI限制位点之间的片段,且克隆到pMON82554穿梭载体的NcoI和BsiWI位点之间。如上所述这些载体的每一个然后用BamHI进行消化,以产生随后克隆到pMON30098的BamHI位点内的片段,以提供最终质粒载体的剩余遗传元件。包含与GFP融合的这些CTP变体的最终质粒载体和载体的剩余遗传元件连同在其构建中使用的引物一起在表4中列出。
表4.构建用于原生质体转染测定的策略2质粒载体的概括
pMON | CTP::GFP变体 | 引物名称 | SEQ ID NO: |
pMON78832 | At-CTP2(C/M)::AgroAS(F298W)::GFP | CTP2-N1-1 | 85 |
CTP2-N1-2 | 86 | ||
CTP2-N1-3 | 87 | ||
CTP2-N1-4 | 88 | ||
CTP2-N1-5 | 89 | ||
CTP2-N1-6 | 90 | ||
CTP2-N1-7 | 91 | ||
CTP2-N1-8 | 92 | ||
CTP2-N1-9 | 93 | ||
CTP2-N1-10 | 94 | ||
CTP2-N1-11 | 95 | ||
CTP2-N1-12 | 96 |
pMON | CTP::GFP变体 | 引物名称 | SEQ ID NO: |
CTP2-N1-13 | 97 | ||
CTP2-N1-14 | 98 | ||
pMON78833 | At-CTP2(E/K)::AgroAS(F298W)::GFP | CTP2-N1-1 | 85 |
CTP2-N1-2 | 86 | ||
CTP2-N1-3 | 87 | ||
CTP2-N1-4 | 88 | ||
CTP2-N1-5 | 89 | ||
CTP2-N2-6 | 99 | ||
CTP2-N1-7 | 91 | ||
CTP2-N1-8 | 92 | ||
CTP2-N2-9 | 100 | ||
CTP2-N1-10 | 94 | ||
CTP2-N1-11 | 95 | ||
CTP2-N1-12 | 96 | ||
CTP2-N1-13 | 97 | ||
CTP2-N1-14 | 98 | ||
pMON78834 | At-CTP2+10::AgroAS(F298W)::GFP | CTP2-N1-1 | 85 |
CTP2-N1-2 | 86 | ||
CTP2-N1-3 | 87 | ||
CTP2-N1-4 | 88 | ||
CTP2-N1-5 | 89 | ||
CTP2-N3-6A | 101 | ||
CTP2-N3-6B | 102 | ||
CTP2-N1-7 | 91 | ||
CTP2-N1-8 | 92 | ||
CTP2-N3-9A | 103 | ||
CTP2-N3-9B | 104 | ||
CTP2-N1-10 | 94 | ||
CTP2-N1-11 | 95 | ||
CTP2-N1-12 | 96 | ||
CTP2-N1-13 | 97 | ||
CTP2-N1-14 | 98 | ||
pMON78835 | At-CTP2+5::AgroAS(F298W)::GFP | CTP2-N1-1 | 85 |
CTP2-N1-2 | 86 | ||
CTP2-N1-3 | 87 | ||
CTP2-N1-4 | 88 | ||
CTP2-N1-5 | 89 | ||
CTP2-N4-6 | 105 | ||
CTP2-N1-7 | 91 | ||
CTP2-N1-8 | 92 | ||
CTP2-N4-9 | 106 |
pMON | CTP::GFP变体 | 引物名称 | SEQ ID NO: |
CTP2-N1-10 | 94 | ||
CTP2-N1-11 | 95 | ||
CTP2-N1-12 | 96 | ||
CTP2-N1-13 | 97 | ||
CTP2-N1-14 | 98 | ||
pMON78138 | Zm-ASA1-CTP::AgroAS(F298W)::GFP | ASA1-1-1T | 107 |
ASA1-1-2T | 108 | ||
ASA1-1-3T | 109 | ||
CTP2-N1-7 | 91 | ||
ASA1-1-5B | 110 | ||
ASA1-1-4B | 111 | ||
ASA1-1-3B | 112 | ||
ASA1-1-2B | 113 | ||
CTP2-N1-8 | 92 | ||
pMON78141 | Zm-ASA1-CTP+20::AgroAS(F298W)::GFP | ASA1-1-1T | 107 |
ASA1-1-2T | 108 | ||
ASA1-20-3T | 114 | ||
ASA1-20-4T | 115 | ||
CTP2-N1-7 | 91 | ||
ASA1-1-5B | 110 | ||
ASA1-1-4B | 111 | ||
ASA1-1-3B | 112 | ||
ASA1-20-3B | 116 | ||
ASA1-20-2B | 117 | ||
CTP2-N1-8 | 92 | ||
pMON69760 | Zm-ASA2-CTP::AgroAS(F298W)::GFP | AS2-5′-1 | 118 |
AS2-5′-41 | 119 | ||
AS2-5′-81 | 120 | ||
AS2-5′-121 | 121 | ||
AS2-5′-161 | 122 | ||
AS2-3′-208 | 123 | ||
AS2-3′-168 | 124 | ||
AS2-3′-128 | 125 | ||
AS2-3′-88 | 126 | ||
AS2-3′-48 | 127 | ||
pMON69761 | Zm-ASA2-CTP+5aa::AgroAS(F298W)::GFP | AS2-5′-161-5aa | 128 |
AS2-3′-183-5aa | 129 |
pMON | CTP::GFP变体 | 引物名称 | SEQ ID NO: |
AS2-3′-153-5aa | 130 | ||
pMON69762 | Zm-ASA2-CTP+10aa::AgroAS(F298W)::GFP | AS2-5′-161-10aa | 131 |
AS2-5′-196-10aa | 132 | ||
AS2-3′-198-10aa | 133 | ||
AS2-3′-163-10aa | 134 | ||
pMON69771 | Zm-ASA2-CTP::GFP | AS2GFP-5′ | 135 |
AS2GFP-3′ | 136 | ||
pMON78135 | Os-Wx-CTP::AgroAS(F298W)::GFP | RW-CM-1T | 137 |
RW-CM-2T | 138 | ||
RW-CM-3T | 139 | ||
RW-CM-4T | 140 | ||
RW-CM-5T | 141 | ||
RW-CM-6T | 142 | ||
RW-CM-7T | 143 | ||
RW-CM-8B | 144 | ||
RW-CM-7B | 145 | ||
RW-CM-6B | 146 | ||
RW-CM-5B | 147 | ||
RW-CM-4B | 148 | ||
RW-CM-3B | 149 | ||
RW-CM-2B | 150 | ||
RW-CM-1B | 151 | ||
pMON78136 | Os-Wx-CTP+5::AgroAS(F298W)::GFP | RW-CM-1T | 137 |
RW-CM-2T | 138 | ||
RW-CM-3T | 139 | ||
RW-CM-4T | 140 | ||
RW-CM-5T | 141 | ||
RW-CM-6T | 142 | ||
RW-5-7T | 152 | ||
RW-5-8T | 153 | ||
RW-CM-8B | 144 | ||
RW-CM-7B | 145 | ||
RW-CM-6B | 146 | ||
RW-CM-5B | 147 | ||
RW-CM-4B | 148 | ||
RW-CM-3B | 149 | ||
RW-5-2B | 154 | ||
RW-5-1B | 155 | ||
pMON | CTP::GFP变体 | 引物名称 | SEQ ID NO: |
pMON78137 | Os-Wx-CTP+20::AgroAS(F298W)::GFP | RW-CM-1T | 137 |
RW-CM-2T | 138 | ||
RW-CM-3T | 139 | ||
RW-CM-4T | 140 | ||
RW-CM-5T | 141 | ||
RW-CM-6T | 142 | ||
RW-20-7T | 156 | ||
RW-20-8T | 157 | ||
RW-20-9T | 158 | ||
RW-CM-8B | 144 | ||
RW-CM-7B | 145 | ||
RW-CM-6B | 146 | ||
RW-CM-5B | 147 | ||
RW-CM-4B | 148 | ||
RW-CM-3B | 149 | ||
RW-5-2B | 154 | ||
RW-20-2B | 159 | ||
RW-20-1B | 160 | ||
pMON69758 | Zm-DHDPS-CTP::AgroAS(F298W)::GFP | DHDPS-5′-1 | 161 |
DHDPS-5′-41 | 162 | ||
DHDPS-5′-81 | 163 | ||
DHDPS-5′-121 | 164 | ||
DHDPS-5′-161 | 165 | ||
DHDPS-3′-AgroAS | 166 | ||
DHDPS-3′-184 | 167 | ||
DHDPS-3′-140 | 168 | ||
DHDPS-3′-99 | 169 | ||
DHDPS-3′-59 | 170 | ||
pMON69759 | Zm-DHDPS-CTP+9aa::AgroAS(F298W)::GFP | DHDPS-5′-161-9aa | 171 |
DHDPS-5′-201-9aa | 172 | ||
DHDPS-3′-216-9aa | 173 | ||
DHDPS-3′-176-9aa | 174 | ||
pMON69765 | Zm-DHDPS-CTP+20aa::AgroAS(F298W)::GFP | DHDPS-5′-201-20aa | 175 |
DHDPS-5′-241-20aa | 176 | ||
DHDPS-3′-249-20aa | 177 | ||
DHDPS-3′-209- | 178 |
pMON | CTP::GFP变体 | 引物名称 | SEQ ID NO: |
20aa | |||
DHDPS-3′-169-20aa | 179 | ||
pMON69764 | Zm-DHDPS-CTP+5aa::AgroAS(F298W)::GFP | DHDPS-5′-161-5aa | 180 |
DHDPS-5′-201-5aa | 181 | ||
DHDPS-3′-244-5aa | 182 | ||
DHDPS-3′-204-5aa | 183 | ||
DHDPS-3′-164-5aa | 184 | ||
pMON69772 | Zm-DHDPS-CTP::GFP | DHDPSGFP-5′ | 185 |
DHDPSGFP-3′ | 186 | ||
pMON69766 | Zm-DHDPS-CTP+5::GFP | DHDPS-N1 | 187 |
DHDPS-N2 | 188 | ||
DHDPS-N3 | 189 | ||
DHDPS-N4 | 190 | ||
DHDPS-N9 | 191 | ||
DHDPS-N10 | 192 | ||
DHDPS-N11 | 193 | ||
DHDPS-N12 | 194 | ||
DHDPS-N13 | 195 | ||
DHDPS-N14 | 196 | ||
DHDPS-N15 | 197 | ||
DHDPS-N18 | 198 |
实施例4
AgroAS和变体至质体的定位
这个实施例描述了用于预测不同叶绿体转运肽(CTPs)在AS::GFP融合蛋白的质体靶向中的能力的2种瞬时表达测定系统。这些测定利用包含邻氨基苯甲酸合酶-绿色荧光蛋白(AS::GFP)融合物和实施例3中描述的融合物的原生质体转染载体。
开发了2种瞬时表达测定法以预测AS::GFP融合蛋白在玉蜀黍细胞中的定位。中等通量玉蜀黍原生质体系统用于就其使AS::GFP融合蛋白靶向变白的玉蜀黍叶细胞的质体的能力筛选许多不同的CTPs。在发育中的玉蜀黍胚中表达蛋白质的较低通量系统用于证实原生质体系统中可见的质体定位模式。来自这2种测定的数据随后用于预测各种CTPs在指导土壤杆菌属和中华根瘤菌属AS蛋白质在转基因玉蜀黍胚中的定位中的能力。
瞬时测定系统中测试的所有构建体用常见的载体主链中相同的遗传元件进行构建,所述常见的载体主链使用e35S启动子、hsp70(DnaK)内含子和nos 3’UTR表达每种基因。
为了确保瞬时测定系统正确发挥作用,构建了对照质粒。关于胞质定位的对照是包含无CTP的GFP融合物(pMON30098;表2和5)、与GFP的AgroAS融合物(不添加CTP)(pMON78818;表2和5)和缺乏Zm-ASA2-CTP的截短的Zm-ASA2-CTP::GFP融合物(pMON78822;表2和5)的载体。用于质体定位的对照是与GFP融合的玉蜀黍ASA2(pMON78820;表2和5)和与GFP融合的At-CTP2(pMON53173;表5)。2种另外的对照用于证实来自转基因植物的数据;CTP1::GFP(pMON79960;表2和5)和CTP1::AgroAS(F298W)::GFP(pMON79961;表2和5)。来自这些载体中每一种的GFP或GFP融合蛋白的定位模式在表5中报告。该数据还证实如实施例1中描述的细胞分级分离结果,指出CTP1::AgroAS(F298W)::GFP融合蛋白的胞质定位和CTP1不能使AgroAS(F298W)靶向质体。
在大多数实验中,对于测试的每种CTP构建另外的对照。这些对照由与GFP直接融合的测试的CTP组成。例如,Zm-ASA2-CTP与GFP融合(pMON69771)以测试Zm-ASA2::AgroAS::GFP融合物。用于构建测试的所有载体的方法在实施例3中详细描述。
对于测试的每一种CTPs进行几个变化。这些变化通过改变对天然宿主蛋白质的CTP添加的N末端氨基酸的数目来区分(表1)。例如,在Zm-ASA2-CTP系列中,使用与GFP直接融合的实验测定的CTP,以及包括来自Zm-ASA2的N末端的5和18个氨基酸的2种另外形式,所述氨基酸从与CTP相邻的成熟的玉蜀黍邻氨基苯甲酸合酶α2亚基的氨基末端的区域中分离。构建体在表1中命名为Zm-ASA2-CTP+5(SEQ ID NO:8)和Zm-ASA2-CTP+18(SEQ ID NO:9)。
作为初步测试,表1中描述的CTP构建体在变白的原生质体系统中进行评估。使用本领域众所周知的方法由变白的玉蜀黍幼苗制备叶肉原生质体(参见例如Sheen,1993)。
使用本领域众所周知的方法将不同的载体电穿孔到原生质体内。约18-24小时后,使用共焦显微镜术就GFP荧光计数原生质体细胞。简言之,使用配备氩离子激光器、绿色和红色氦-氖激光器以及变色二极管激发的激光器(Coherent Laser Division,Santa Clara,CA)的Zeiss激光器扫描显微镜(Zeiss Laser Scanning Microscope)LSM510 META(Carl ZeissMicroImaging,Inc.,Thornwood,NY)进行显微镜术。使用LSM 5 ImageExaminer,版本3.2.0.70(Carl Zeiss MicroImaging,Inc.,Thornwood,NY)进行图像采集和分析。使用Adobe Photoshop CS,版本8.0(AdobeSystems Incorporated,San Jose,CA)进行图像处理。就在下述中具有GFP荧光而言评分关于每种构建体的至少50个细胞;a)仅质体,b)仅细胞溶胶和c)质体和细胞溶胶。
作为次级测试,开发了特异性瞬时测定系统以评估设计为在发育中的玉蜀黍胚组织中表达的构建体。基于在包含仅质体定位的50%或更多转化细胞的原生质体系统中具有得分来选择通过这种测试评估的构建体。还测试了如上详述的另外的对照构建体。
在授粉后约14天(DAP)时,从来自玉蜀黍例如HiII或近交亲本的表面灭菌谷穗中分离胚。将约20-30个胚置于包含补加有0.2M山梨糖醇+0.2M甘露糖醇的N6培养基的Petri板上(Chu,1978)。在室温下温育4小时后,使用BiolisticTM PDS-1000/He粒子递送系统(Particle DeliverySystem)(Bio-Rad Corporation)用由DNA包被的0.6μm金颗粒轰击胚。板在从停止屏到靶架9cm处轰击2次,具有1100psi的破裂压和1cm的间隙距离。轰击后,在如前所述通过多光子共焦显微镜术分析前使胚于28℃在黑暗中温育过夜。每种样品计数至少50个细胞且如上文关于原生质体系统所述进行评分。
来自定位测定的结果显示于表5中。结果指出基于显现,下述6种CTPs使AgroAS有效靶向玉蜀黍生殖细胞中的质体:Zm-ASA2-CTP+18(SEQ ID NO:9;pMON78824的CTP组分)、Rg-AS长-CTP(SEQ ID NO:14;pMON78142的CTP组分)、Rg-AS短-CTP(SEQ ID NO:13;pMON78143和pMON78139的CTP组分)、At-CTP2(E/K)(SEQ ID NO:2;pMON78833的CTP组分)、At-CTP2(E/K)+10(SEQ ID NO:3;pMON78834的CTP组分)和Zm-DHDPS-CTP+20(SEQ ID NO:17;pMON69765的CTP组分)。结果还指出并非所有测定的各种CTPs在原生质体或胚瞬时表达测定中就定位AS::GFP的能力都是阳性的。这些结果确证了实施例1中描述的结果,且指出鉴定关于使单体AS蛋白质成功定位在单子叶植物的叶绿体中且因此产生升高的色氨酸水平的有用CTPs的需要。
表5.定位测定的结果
pMONID | CTP::AgroAS::GFP变体 | 原生质体%质体定位 | 胚%质体定位 |
78832 | At-CTP2(C/M)::AgroAS(F298W)::GFP | 68.6 | 未测试 |
78833 | At-CTP2(E/K)::AgroAS(F298W)::GFP | 64 | 79 |
78834 | At-CTP2+10::AgroAS(F298W)::GFP | 72 | 35 |
78835 | At-CTP2+5::AgroAS(F298W)::GFP | 60.6 | 未测试 |
53173 | 对照:At-CTP2::GFP | >80 | 80 |
78138 | Zm-ASA1-CTP::AgroAS(F298W)::GFP | 0 | 未测试 |
78141 | Zm-ASA1-CTP+20::AgroAS(F298W)::GFP | 0 | 未测试 |
69760 | Zm-ASA2-CTP::AgroAS(F298W)::GFP | 0 | 未测试 |
69761 | Zm-ASA2-CTP+5::AgroAS(F298W)::GFP | 0 | 未测试 |
78824 | Zm-ASA2-CTP+18::AgroAS(wt)::GFP | 100 | 81 |
69771 | 对照:Zm-ASA2-CTP::GFP | 15 | 未测试 |
78135 | Os-Wx-CTP::AgroAS(F298W)::GFP | 0 | 未测试 |
78136 | Os-Wx CTP+5::AgroAS(F298W)::GFP | 0 | 未测试 |
78137 | Os-Wx-CTP+20::AgroAS(F298W)::GFP | 72 | 未测试 |
78140 | 对照:Os-Wx-CTP::GFP | 77 | 未测试 |
78143 | Rg-AS 短-CTP::AgroAS(F298W)::GFP | 873 | 54 |
78142 | Rg-AS 长-CTP::AgroAS(F298W)::GFP | 88.6 | 55 |
78139 | 对照:Rg-AS 短-CTP::GFP | 77.3 | 67 |
69758 | Zm-DHDPS-CTP::AgroAS(F298W)::GFP | 0 | 未测试 |
69759 | Zm-DHDPS-CTP+9::AgroAS(F298W)::GFP | 3 | 未测试 |
69765 | Zm-DHDPS-CTP+20::AgroAS(F298W)::GFP | 71 | 在质体和细胞溶胶中91%;9%仅在细胞溶胶中 |
69763 | Zm-DHDPS-CTP+3::AgroAS(F298W)::GFP | 0 | 33 |
69772 | 对照:Zm-DHDPS-CTP::GFP | 0 | 10 |
69766 | 对照:Zm-DHDPS-CTP+5::GFP | 30 | 未测试 |
69774 | At-CTP2(E/K)::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 64 | 未测试 |
69775 | At-CTP2+10::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 72.7 | 未测试 |
69776 | Zm-ASA2-CTP::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 0 | 未测试 |
69777 | Zm-ASA2-CTP+18::苜蓿根瘤菌邻氨基苯甲酸合酶::GFP | 70.7 | 未测试 |
78818 | 对照:无CTP::AgroAS(F298W)::GFP | 0 | 0 |
78820 | 对照:Zm-ASA2(包括CTP)::GFP | 70.3 | 85 |
pMONID | CTP::AgroAS::GFP变体 | 原生质体%质体定位 | 胚%质体定位 |
78822 | 对照:无CTP::成熟的Zm-ASA2::GFP | * | 未测试 |
79961 | 对照:CTP1::AgroAS(F298W)::GFP | * | 0 |
30098 | 对照:GFP | * | 0 |
79960 | 对照:CTP1::GFP | ** | 44 |
*对于这些对照,在原生质体瞬时测定中未观察到质体定位;数据未定量。
**对于这种对照,观察到质体和胞质定位但未定量。
实施例5
转化载体和玉蜀黍转化
这个实施例描述了包含核酸序列的转化载体的构建,所述核酸序列编码与各种叶绿体转运蛋白组合的根癌土壤杆菌和苜蓿根瘤菌邻氨基苯甲酸合酶(AS)的野生型(wt)和突变等位基因。这个实施例还提供了用于由本文描述的转化载体转化玉蜀黍的规程。
对于包含野生型根癌土壤杆菌AS的转化载体的构建,通过用XhoI消化、使用绿豆核酸酶使末端平端化且随后用BglII切割以分离第一个片段,从质粒pMON66580中切割出AS编码序列。为了分离包含玉蜀黍油质蛋白启动子、hsp70内含子和Tr7-3’UTR的第二个片段,用BglII和SmaI切割质粒pMON69753。将这2个片段连接以产生pMON69754。为了构建包含Zm-ASA2-CTP的最终转化载体,用XhoI消化pMON69754,且分离包含玉蜀黍油质蛋白启动子、hsp70内含子、与-wt融合的Zm-ASA2-CTP和Tr7-3’UTR的纯化片段。随后将这个片段连接到载体pMON78808中,所述载体pMON78808也已用XhoI进行消化以产生最终转化载体pMON69755(图6)。
对于包含AgroAS(F298W)突变等位基因的转化载体的构建,pMON66869用StuI和RsrII进行消化,以分离包含部分AgroAS(F298W)编码区的片段。类似地,pMON69754也用StuI和RsrII进行消化,以分离包含玉蜀黍油质蛋白启动子、hsp70内含子、部分编码区和Tr7-3’-UTR的片段。使所得到的片段连接以产生pMON69756。为了构建最终转化载体,用XhoI消化pMON69756,且纯化包含玉蜀黍油质蛋白启动子、hsp70内含子、与AgroAS(F298W)融合的Zm-ASA2-CTP和Tr7-3’UTR的片段。随后将这个片段连接到pMON78808的XhoI位点内,以产生pMON69757;SEQ ID NO:215)。
类似地构建包含AgroAS(S51F)编码区(SEQ ID NO:203)的转化载体。例如,上文描述的质粒pMON69754用BamHI和NcoI进行消化,以取出AgroAS编码区。质粒pMON58121也用BamHI和NcoI进行消化,以分离AgroAS(S51F)突变等位基因。随后使所得到的片段连接以产生中间质粒pMON69767。如上所述通过用XhoI消化pMON69767且在pMON78808的XhoI位点处连接片段来构建最终转化载体pMON69768。
包含AgroAS(S51C)等位基因的转化载体的构建的例子是pMON68065。pMON68063用XhoI限制酶进行消化,在1.0%琼脂糖凝胶上进行分离,且从凝胶分离对应于与Zmhsp70(Zm.DNAK)内含子、AgroAS(S51C)等位基因和Os-gt1-3’UTR融合的玉蜀黍油质蛋白启动子的4569bp DNA片段。
pMON78808 DNA也用XhoI限制酶进行切割,使用碱性磷酸酶(NewEngland BioLabs Inc.)进行去磷酸化,并且使用QIAGEN PCR纯化试剂盒(QIAGEN Inc.)纯化DNA。使pMON78808的XhoI消化的DNA和pMON68063的4569bp XhoI消化的DNA片段连接在一起以产生pMON68065。
包含AgroAS(S51C)密码子最优化的(AgroAS(S51C)-nno)等位基因的转化载体的构建的例子是pMON68066。AgroAS(S51C)突变等位基因的密码子最优化如美国专利申请系列号11/503,532中描述的进行,所述专利引入本文作为参考)。密码子最优化的AgroAS(S51C)-nnoDNA通过BLUE HERON BIOTECHNOLOGY组(Bothell,WA,USA)进行合成。合成DNA随后用NcoI和BamHI限制酶进行消化,在1.0%琼脂糖凝胶上进行分离,并且如上所述从凝胶切割出对应于AgroAS(S51C)-nno的2193bp DNA片段且进行纯化。pMON68063的DNA也用NcoI和BamHI限制酶进行切割且在1.0%琼脂糖凝胶上进行分离。分离的5244bp NcoI/BamHI消化的DNA片段包含pMON68063中描述的所有遗传元件,除了AgroAS(S51C)突变等位基因。连接反应通过使NcoI/BamHI消化的AgroAS(S51C)-nno DNA(2193bp片段)和pMON68063(5244bp DNA片段)的DNA混合来进行,以产生pMON68064。pMON68064随后用XhoI限制酶进行切割,在1.0%琼脂糖凝胶上进行分离,且如上所述纯化包含与hsp70(Zm.DNAK)内含子、AgroAS(S51C)-nno和Os-gt1-3’UTR融合的玉蜀黍油质蛋白启动子的4532bp DNA片段。pMON78808 DNA的DNA也用XhoI限制酶进行切割,使用碱性磷酸酶(New England BioLabs Inc.,Beverly,MA)进行去磷酸化,并且使用QIAGEN PCR纯化试剂盒(QIAGEN Inc.,Valencia,CA)进行纯化。连接反应通过使去磷酸化的XhoI-切割的pMON78808 DNA和pMON68064的4532bp bp DNA片段混合来进行,以产生pMON68066。
使用类似克隆策略将质粒pMON69769用作基本载体以产生各种其他转化载体,包括pMON82561(图7)、pMON78152(图8)、pMON78153(图9)、pMON82560(图10)、pMON69779(图11)、pMON78846、pMON78850、pMON78851、pMON68065、pMON69781、pMON94548、pMON94549、pMON97701、pMON97703和pMON97705。
玉蜀黍的转化
基本上如美国专利申请公开20050005327中所述将上文描述的转化载体转化到玉蜀黍内,所述专利申请整体引入本文作为参考。简言之,在授粉后约10天收获包含未成熟的胚的谷穗且于4℃保持冷冻直至使用(最多至收获后5天)。关于这种转化方法的优选胚大小是~1.5-2.0mm。这个大小通常在授粉后10天在温室内达到,其中生长条件为平均温度87°F、日长14小时伴随由GE 1000瓦高压钠灯提供的补充照明。
从表面灭菌的谷穗分离未成熟的胚且直接投入在1.5-mL微量离心机管中制备的土壤杆菌属细胞悬浮液中。分离连续持续约5-60分钟。可替代地,将胚直接切到接种培养基(不含土壤杆菌属或乙酰丁香酮)内进行5-60分钟,且随后用土壤杆菌属细胞悬浮液接种5-30分钟。使用精细包尖头(tipped)的无菌转移移液管取出土壤杆菌属细胞悬浮液后,将未成熟的胚转移到crn398共培养培养基上(表6)。随后将胚置于培养基上,其中小盾片侧朝上。胚在暗培养箱(23℃)中培养约14-48小时。
随后将胚转移到愈伤组织诱导培养基(crn336,表6)上,所述愈伤组织诱导培养基包含0.1mM草甘膦和500mg/L羧苄青霉素,以抑制培养皿(100mm x 25mm)中的土壤杆菌属。培养物在暗培养室/培养箱中于30℃温育2周,随后在暗培养室/培养箱中于27℃再温育1周。随后将所有愈伤组织碎片个别转移到第一种再生培养基(crn335,表6)上,所述第一种再生培养基包含0.1mM草甘膦和250mg/L羧苄青霉素。培养物在这种培养基上在具有16小时光/8小时暗光周期的27℃培养室中生长7-10天。随后将它们转移到培养皿(100mm x 25mm)中的第二种再生培养基(crn333,表6)上,于27℃伴随16小时光约2-3周。然后将具有再生枝条和活组织的所有愈伤组织碎片转移到新鲜的crn333板或crn334PHYTATRAYs(表6)上或直接转移至包含生根培养基(crn366,表6)的圣代(sundae)杯,以在转移至土壤之前进一步生长(约1-3周)。再生培养基(crn335、crn333和crn334)都包含250mg/L羧苄青霉素和0.1mM草甘膦。
然后将小植株转移至土壤,在生长室中在27℃、80%湿度和低光强度下使其茁壮(harden off)约1-2周,然后转移至温室且在标准温室条件下生长。收集所得到的仁且如下所述进行分析。
表6.在玉米转化中使用的培养基的组成(每升)。
组分 | 共培养培养基(crn398) | 愈伤组织诱导培养基(crn336) | 第一种再生培养基(crn335) | 第二种再生培养基(crn333(板)/crn334(PHYTATRAYs)) | 生根培养基(crn366) |
MS盐 | 2.17g | 4.33g | 4.33g | 4.33g | 2.17g |
蔗糖 | 20g | 30g | 30g | 20g | |
麦芽糖 | 20g | ||||
葡萄糖 | 10g | 10g | |||
l-脯氨酸 | 115mg | 1.38g | 1.38g | ||
酪蛋白氨基酸 | 0.5g | 0.05g | |||
IBA(1mg/mL原液) | 0.75mL | ||||
l-天冬酰胺 | 0.15g | ||||
肌醇 | 0.1g | ||||
NAA(1mg/mL原液) | 0.5mL | ||||
硫胺素-HCl(0.5mg/mL原液) | 1.0mL | 1.0mL | |||
2,4-D(1mg/mL原液) | 3.0mL | 0.5mL | |||
硝酸银(2mg/mL原液) | 1.7mL | 1.7mL | |||
MS维生素100X | 10mL | 10mL | 5.0mL | ||
MS Fromm1000X | 1.0mL | 1.0mL | |||
羧苄青霉素(250mg/mL) | 2.0mL | 1.0mL | 1.0mL | ||
草甘膦(0.5M原液) | 0.2mL | 0.2mL | 0.2mL | 0.2mL | |
Phytagel | 3.0g | 3.0g | 3.0g | 3.0g | |
BAP(0.5mg/mL原液) | 0.02mL | 7.0mL | |||
乙酰丁香酮(1.0M) | 0.2mL | ||||
琼脂糖低EEO | 5.5g |
实施例6
用于玉蜀黍仁中的AS的免疫定位和色氨酸的定量的方法
这个实施例描述了在来自用实施例5中描述的CTP构建体转化的玉蜀黍事件的仁中的免疫定位研究和游离色氨酸水平的定量中使用的分析程序。
在26 DAP时分开收获来自具有不同构建体的F1阶段转基因植物的仁。这些构建体包括pMON69755(图6)、pMON69779(图11)、pMON82560(图10)、pMON82561(图7)、pMON78153(图9)和pMON78152(图8)。从这些构建体的每个转基因系中分离8-10个胚,在3.7%甲醛溶液中固定,且在通过下文描述的免疫标记研究就表达蛋白质的定位进行分析前贮存于4℃。
还使用DNeasyTM植物试剂盒(目录号69104,Qiagen,Waltham,MA)从这些转基因仁的胚乳提取基因组DNA。使用在序列表中分别命名为SEQ ID NO:226和SEQ ID NO:227的寡核苷酸引物玉蜀黍AS2-5′和玉蜀黍AS2-3′进行PCR扩增以鉴定阳性和阴性仁。
上文描述的玉米胚在500μl 0.05%Triton X-100中温育30分钟。使用配备蓝宝石(sapphire)刀的Leica VT 1000S振动刀片切片机将小盾片组织切片切片成20μm厚的切片,然后在包含10mM甘氨酸的PBS缓冲液(137mM NaCl、3mM KCl、8mM Na2HPO4和1.5mM KH2PO4)中洗涤3次,每次洗涤10分钟。
组织切片随后在酶促缓冲液(4%果胶酶、2%纤维素酶)中于28℃温育40分钟。组织然后用未稀释的血清封闭15分钟,使用山羊血清(Sigma目录号G-9023;Sigma Chemical Company,St.Louis,MO)用于AgroAS检测和兔血清(Sigma R-9133)用于玉蜀黍AS。组织切片然后在溶于PBS缓冲液的1%兔抗his AgroAS或山羊抗玉蜀黍AS中于4℃伴随轻轻摇动温育过夜。
过夜温育后,组织在包含10mM甘氨酸的PBS缓冲液中洗涤20分钟,且在分别的未稀释血清中在室温下再次温育15分钟。组织然后在黑暗中与Alexa缀合物第二抗体在室温下温育2小时。稀释因子对于用于AgroAS蛋白质的532(Molecular Probes,Inc.,Eugene,OR)山羊抗兔缀合物第二体抗为1∶1000,和对于用于Zm AS蛋白质的488(Molecular Probes,Inc.)兔抗山羊为1∶1000。温育后,组织在包含10mM甘氨酸的PBS中洗涤3次,每次洗涤10分钟。组织然后用对于细胞壁(Sigma)溶液(在原液中2.5mg/ml,用蒸馏水1∶50稀释)的卡尔科弗卢尔(calcofluor)和对于核的Hoechst染料(Molecular Probes Inc.)在黑暗中伴随摇动复染15分钟。染色的组织随后在封固前在蒸馏水中洗涤10分钟。
结果指出当样品用具有合并通道的AgroAS(在543nm处的红色荧光)和对照Zm AS抗体(在488nm处的绿色荧光)进行探测时,黄色的出现暗示蛋白质在质体中的共定位(数据未显示)。
为了提取氨基酸级分,将30mg磨碎的仁置于离心机小瓶中。向每种样品中加入1毫升5%三氯乙酸。样品通过涡旋进行混合,且置于4℃过夜。样品然后再次混合且在微量离心机中在14,000rpm下旋转15分钟。随后取出一些上清液,置于HPLC小瓶中且密封。样品在分析前保存于4℃。
如Agilent技术出版物,“Amino Acid Analysis Using ZorbaxEclipse-AAA Columns and the Agilent 1100HPLC”,2000年3月17日中所述进行氨基酸分析。该分析用邻苯二甲醛(o-pthaldialdehyde)(OPA)预衍生氨基酸,然后通过反相层析分离OPA缀合物。分离使用Agilent1100系列HPLC系统(Agilent,Palo Alto,CA)来进行,其使用EclipseXDB-C185μm、4.6x 150mm柱和1.2ml/分钟的流速。氨基酸浓度使用荧光进行测量:在340nm处激发、在450nm处发射。洗脱用根据表7的HPLC缓冲液A和B的梯度,其中HPLC缓冲液A是40mM Na2HPO4,pH7.8且HPLC缓冲液B是9∶9∶2::甲醇∶乙腈∶水。
表7:氨基酸洗脱
时间 | 0 | 20 | 21 | 26 | 27 |
%缓冲液A | 95 | 35 | 0 | 0 | 0 |
%缓冲液B | 5 | 65 | 100 | 100 | 100 |
制备或购买浓度范围为0-100μg/ml的氨基酸标准。脯氨酸分析需要使用9-芴基甲基氯甲酸酯(FMOC)的另外衍生步骤。如表8中所示的结果以ppm报告。
实施例7
玉蜀黍中的游离色氨酸水平的分析
这个实施例阐述了用实施例5中描述的表达载体转化的玉蜀黍植物中的游离色氨酸水平的分析。
F1仁如实施例6中所述就游离色氨酸进行分析。下表8中所示的来自一些转化的植物的结果指出在瞬时表达测定中证实成功使GFP靶向的能力(表5中所示的数据)的CTPs在转化的植物中具有更高水平的游离色氨酸。此外,未证实使GFP定位在质体中的能力的CTPs具有较低水平的游离色氨酸。这些结果指出并非所有CTPs都可以指导单体AS定位至质体且因此增加游离色氨酸水平。
表8.F3纯合转基因玉蜀黍植物中的平均游离色氨酸水平。
构建体 | 载体描述 | 事件ID | 色氨酸的平均值ppm | 色氨酸的StdDevppm |
PMON30167 | 仅载体 | RAG120 | 11 | 1 |
亲本系 | 非转基因的 | 11 | 2 | |
PMON69755 | Zm-ASA2-CTP+18:_wt AS | ZM_S100593 | 118 | 8 |
ZM_S101241 | 99 | 45 | ||
ZM_S103021 | 58 | 18 | ||
ZM_S103022 | 63 | 6 | ||
ZM_S98891 | 122 | 26 | ||
PMON69757 | Zm-ASA2-CTP+18:_F298W | ZM_S100715 | 352 | 42 |
ZM_S101226 | 310 | 16 | ||
ZM_S99658 | 286 | 37 |
构建体 | 载体描述 | 事件ID | 色氨酸的平均值ppm | 色氨酸的StdDevppm |
ZM_S99679 | 242 | 16 | ||
ZM_S99787 | 319 | 22 | ||
PMON69768 | Zm-ASA2-CTP+18:_S51F | ZM_S104037 | 227 | 18 |
ZM_S104038 | 129 | 11 | ||
ZM_S104052 | 174 | 18 | ||
ZM_S105091 | 258 | 30 | ||
ZM_S105105 | 254 | 53 | ||
PMON69770 | Zm-ASA2-CTP+18:_S51C | ZM_S102618 | 677 | 174 |
ZM_S102637 | 649 | 84 | ||
ZM_S103233 | 585 | 190 | ||
ZM_S103234 | 612 | 197 | ||
ZM_S103245 | 632 | 160 | ||
ZM_S103257 | 667 | 106 | ||
ZM_S103261 | 430 | 106 | ||
ZM_S103277 | 684 | 251 | ||
PMON69779 | ZM-DHDPS-CTP+20:_F298W | ZM_S115297 | 150 | 16 |
ZM_S115309 | 129 | 32 | ||
ZM_S115313 | 123 | 30 | ||
ZM_S115314 | 148 | 15 | ||
ZM_S116346 | 145 | 12 | ||
PMON67146 | Zm-DHDPS-CTP+3:_F298W | ZM_S67192 | 49 | 47 |
ZM_S67201 | 41 | 49 | ||
ZM_S67209 | 51 | 48 | ||
PMON78152 | Rg 长-CTP:_F298W | ZM_S111632 | 68 | 12 |
ZM_S112752 | 53 | 13 | ||
ZM_S112777 | 69 | 22 | ||
ZM_S113086 | 89 | 36 | ||
ZM_S113096 | 280 | 43 | ||
PMON78153 | Rg 短-CTP:_F298W | ZM_S113911 | 326 | 14 |
ZM_S113927 | 258 | 99 | ||
ZM_S113946 | 280 | 54 | ||
ZM_S113953 | 241 | 47 | ||
ZM_S114105 | 271 | 79 | ||
PMON82560 | At-CTP2(E/K)+1:_F298W | ZM_S115009 | 353 | 93 |
ZM_S115027 | 340 | 24 | ||
ZM_S115029 | 393 | 25 | ||
ZM_S115066 | 358 | 113 | ||
ZM_S115070 | 361 | 91 | ||
PMON82561 | At-CTP2(E/K)+10:_F298W | ZM_S113026 | 156 | 37 |
ZM_S113029 | 192 | 27 | ||
ZM_S113033 | 147 | 50 | ||
ZM_S113194 | 145 | 40 | ||
ZM_S113833 | 169 | 24 | ||
PMON66559 | CTP1:_F298W | ZM_S44537 | 10 | 2 |
ZM_S44538 | 8 | 1 | ||
ZM_S44540 | 11 | 3 | ||
ZM_S44542 | 8 | 0 | ||
ZM_S46820 | 9 | 2 | ||
ZM_S46828 | 9 | 2 |
构建体 | 载体描述 | 事件ID | 色氨酸的平均值ppm | 色氨酸的StdDevppm |
ZM_S46833 | 10 | 2 | ||
ZM_S46841 | 9 | 1 | ||
ZM_S46842 | 10 | 2 | ||
ZM_S46848 | 11 | 3 | ||
ZM_S46855 | 10 | 2 | ||
ZM_S46859 | 10 | 2 | ||
ZM_S46861 | 9 | 1 | ||
ZM_S46865 | 9 | 1 | ||
ZM_S46873 | 8 | 1 | ||
ZM_S46877 | 9 | 3 | ||
ZM_S46878 | 10 | 1 | ||
PMON68065 | Zm-ASA2-CTP+18:_S51C | ZM_S138234 | 435 | 42 |
ZM_S142738 | 512 | 95 | ||
ZM_S137274 | 514 | 78 | ||
ZM_S139817 | 529 | 108 | ||
ZM_S134462 | 587 | 71 | ||
ZM_S142742 | 594 | 135 | ||
ZM_S142745 | 594 | 90 | ||
ZM_S142725 | 613 | 87 | ||
ZM_S142747 | 651 | 93 | ||
ZM_S142728 | 814 | 395 | ||
PMON68066 | Zm-ASA2-CTP+18:_S51C_nno | ZM_S134079 | 436 | 132 |
ZM_S133073 | 656 | 127 | ||
ZM_S133076 | 774 | 57 | ||
ZM_S134078 | 904 | 119 | ||
ZM_S134080 | 917 | 268 | ||
ZM_S133092 | 928 | 255 | ||
ZM_S133080 | 943 | 154 | ||
ZM_S133098 | 960 | 321 | ||
ZM_S133099 | 1002 | 556 | ||
ZM_S133082 | 1023 | 129 | ||
ZM_S133433 | 1100 | 138 | ||
ZM_S133093 | 1110 | 136 | ||
ZM_S133089 | 1289 | 139 | ||
ZM_S133434 | 1352 | 101 | ||
PMON78850 | At-CTP2(E/K)+1:根瘤菌属_wt | ZM_S127477 | 45 | 4 |
ZM_S129898 | 32 | 6 | ||
ZM_S129905 | 35 | 3 | ||
ZM_S129913 | 45 | 5 | ||
ZM_S129917 | 40 | 7 | ||
PMON78851 | At-CTP2(E/K)+1:根瘤菌属_S51C | ZM_S128450 | 595 | 103 |
ZM_S128451 | 547 | 96 | ||
ZM_S128458 | 983 | 31 | ||
ZM_S128461 | 477 | 85 | ||
ZM_S128463 | 548 | 72 | ||
ZM_S128464 | 547 | 100 | ||
ZM_S128466 | 503 | 39 | ||
ZM_S128467 | 603 | 126 | ||
ZM_S128469 | 532 | 69 |
实施例8
膳食和饲料产物的产生
这个实施例阐述了用于产生包含已在实施例5中描述的表达载体的膳食和饲料产物的方法。
本发明的任何植物或其部分可以进行加工以产生饲料、膳食、蛋白质或油制剂,包括在总色氨酸含量中高的饲料、膳食、蛋白质和油制剂。用于这个目的的特别优选的植物部分是种子。在优选实施方案中,饲料、膳食、蛋白质或油制剂被设计用于牲畜动物或人或两者。产生饲料、膳食、蛋白质和油制剂的方法是本领域已知的。参见,例如,美国专利号4,957,748;5,100,679;5,219,596;5,936,069;6,005,076;6,146,669;和6,156,227。在优选实施方案中,饲料、膳食、蛋白质或油制剂是高色氨酸制剂。此种高色氨酸制剂优选具有超过约200-400ppm、更优选400-600ppm且甚至更优选600-800ppm的色氨酸含量。
实施例9
用于检测CTP::AS转基因的方法
这阐述了可以用于检测在转基因植物细胞、或者衍生自此种转基因植物细胞的饲料或膳食产物中属于包含本文描述的任何CTP::AS序列组合的构建体的独特序列的存在的方法。
CTP::AS DNA的PCR扩增
来自转基因植物细胞、或者来自衍生自此种转基因植物细胞的饲料或膳食产物的基因组DNA可以根据制造商的规程使用QIAGENDNeasyTM Plant mini试剂盒(目录#69104;QIAGEN Inc.)进行提取。PCR反应混合物一般可以包括约100mg提取的基因组DNA、1X PCR反应缓冲液(Expand High Fidelity PCR System,目录#1732641;Roche Inc.,Nutley,NJ)、0.2mM dNTP、5皮摩尔每种引物、3.5单位高保真度Taq聚合酶和水至50μl的终体积。例如,SEQ ID NO:228和SEQ ID NO:229;或SEQ ID NO:230和SEQ ID NO:231的引物对可以用于扩增提取的DNA,以产生对于来自植物细胞的Zm-ASA2+18-CTP::AgroAS构建体的DNA诊断性的扩增子,所述植物细胞已用pMON68066进行转化。对反应混合物实施各种温度循环,其可以包括:于95℃变性、于60℃退火和于72℃延伸,进行35个循环。在35个PCR循环后,约10μl的反应混合物在1.0%琼脂糖凝胶上分离。2649碱基对片段(具有引物对SEQ ID NO:228和SEQ ID NO:229)或2735碱基对片段(具有引物对SEQ ID NO:230和SEQ ID NO:231)的存在指出植物细胞、饲料或膳食产物包含与AS等位基因融合的Zm-ASA2+18-CTP。识别且设计另外的PCR引物对完全在本领域普通技术人员的方法内,所述另外的引物对在检测属于包含本文描述的任何其他CTP::AS序列组合的构建体的独特序列中将是有用的。
如由上文公开内容指导的,无需过度实验即可制备且使用本文公开和要求保护的所有材料和方法。尽管本发明的材料和方法已就优选实施方案和举例说明性实施例而言进行描述,但对于本领域技术人员将显而易见的是,可以在不背离本发明的概念、精神和范围的情况下对本文描述的材料和方法应用变化。对于本领域技术人员显而易见的所有此种类似置换和修饰被视为在如附加权利要求限定的本发明的精神、范围和概念内。
参考文献
下述参考文献特别引入本文作为参考,其程度至它们提供补充本文阐述的那些的示例性程序或其他细节。
美国专利、申请和申请公开:美国申请09/757,089;美国申请10/138,927;美国申请10/430,011;美国申请10/732,721;美国申请11/503,532;美国专利4,957,748;美国专利4,957,748;美国专利5,100,679;美国专利5,100,679;美国专利5,188,642;美国专利5,219,596;美国专利5,219,596;美国专利5,290,924;美国专利5,322,783;美国专利5,322,938;美国专利5,322,938;美国专利5,352,605;美国专利5,359,142;美国专利5,378,619;美国专利5,378,619;美国专利5,424,412;美国专利5,530,196;美国专利5,538,880;美国专利5,550,318;美国专利5,563,055;美国专利5,610,042;美国专利5,641,876;美国专利5,728,925;美国专利5,837,848;美国专利5,858,741;美国专利5,936,069;美国专利5,936,069;美国专利6,005,076;美国专利6,005,076;美国专利6,118,047;美国专利6,140,078;美国专利6,146,669;美国专利6,146,669;美国专利6,156,227;美国专利6,156,227;美国专利6,175,060;美国专利6,177,611;美国专利6,232,526;美国专利6,252,138;美国专利6,294,714;美国专利6,426,446;美国专利6,429,357;美国专利6,429,362;美国专利6,433,252;美国专利6,437,217;美国专利6,635,806;美国专利号5,641,876;美国专利7,151,204;美国专利号7,217,865;美国公开20030097677;美国公开20030213010;美国公开20050005327
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序列表
<110>MANJUNATH,SIVA
NAVARRO,SANTIAGO XAVIER
RAPP,WILLIAM D.
SHI,XIAOHONG
VARAGONA,MARGUERITE J.
WINSON,JENNIFER L.
YE,GUANGNING
<120>通过邻氨基苯甲酸合酶的叶绿体靶向表达产生高色氨酸玉蜀黍
<130>MONS:110WO
<140>UNKNOWN
<141>2007-08-09
<150>60/837,200
<151>2006-08-11
<160>231
<210>1
<211>228
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>1
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcgtgc 228
<210>2
<211>231
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>2
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcggagaa a 231
<210>3
<211>258
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>3
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcggagaa agcgtcggag 240
attgtacttc aacccatt 258
<210>4
<211>243
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>4
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcggagaa agcgtcggag 240
att 243
<210>5
<211>96
<212>DNA
<213>玉蜀黍(Zea mays)
<400>5
atggccaccg ccagcctcgc gctctcgctg cgcctcgcgc cgtactcgca cccgctgagc 60
ctccgccgcc gcggcgccgc cggcgtcacc tgccgc 96
<210>6
<211>156
<212>DNA
<213>玉蜀黍(Zea mays)
<400>6
atggccaccg ccagcctcgc gctctcgctg cgcctcgcgc cgtactcgca cccgctgagc 60
ctccgccgcc gcggcgccgc cggcgtcacc tgccgcgcca ccaccgccac gttccaccag 120
cttgacgccg tcgcggtgag ggaggaggag tccagg 156
<210>7
<211>142
<212>DNA
<213>玉蜀黍(Zea mays)
<400>7
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc ta 142
<210>8
<211>191
<212>DNA
<213>玉蜀黍(Zea mays)
<400>8
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc tgctgccgtg acgccgatgg taacgatcat tcaggatgac 180
ggagcggaga c 191
<210>9
<211>195
<212>DNA
<213>玉蜀黍(Zea mays)
<400>9
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc tgctgccgtg acgccgcagg cgagcccagt gattagcagg 180
agcgctgcgg cggcc 195
<210>10
<211>227
<212>DNA
<213>稻(0ryza sativa)
<400>10
gcggctctca ccacgtccca gctcgccacc tcggccaccg gcttcggcat cgccgacagg 60
tcggcgccgt cgtcgctgct ccgccacggg ttccagggcc tcaagccccg cagccccgcc 120
ggcggcgacg cgacgtcgct cagcgtgacg accagcgcgc gcgcgacgcc caagcagcag 180
cggtcggtgc agcgtggcag ccggaggttc ccctccgtcg tcgtgtg 227
<210>11
<211>242
<212>DNA
<213>稻(Oryza sativa)
<400>11
gcggctctca ccacgtccca gctcgccacc tcggccaccg gcttcggcat cgccgacagg 60
tcggcgccgt cgtcgctgct ccgccacggg ttccagggcc tcaagccacg cagcccagcc 120
ggcggcgacg cgacgtcgct cagcgtgacg accagcgcgc gcgcgacgcc caagcagcag 180
cggtcggtgc agcgtggcag ccggaggttc ccatccgtcg tcgtgtacgc caccggcgcc 240
gg 242
<210>12
<211>291
<212>DNA
<213>稻(Oryza sativa)
<400>12
atggcggctc tcaccacgtc ccagctcgcc acctcggcca ccggcttcgg catcgccgac 60
aggtcggcgc cgtcgtcgct gctccgccac gggttccagg gcctcaagcc acgcagccca 120
gccggcggcg acgcgacgtc gctcagcgtg acgaccagcg cgcgcgcgac gcccaagcag 180
cagcggtcgg tgcagcgtgg cagccggagg ttcccatccg tcgtcgtgta cgccaccggc 240
gccggcatga acgtcgtgtt cgtcggcgcc gagatggccc cctggagcaa g 291
<210>13
<211>215
<212>DNA
<213>芸香(Ruta graveolens)
<400>13
atgagtgcag cggcaacgtc gatgcaatcc cttaaattct ccaaccgtct ggtcccaccc 60
agtcgccgtc tgtctccggt tccgaacaat gtcacctgca ataacctccc caagtctgca 120
gctcccgtcc ggacagtcaa atgctgcgct tcttcctgga acagtaccat caacggcgcg 180
gccgccacga ccaacggtgc gtccgccgcc agttc 215
<210>14
<211>275
<212>DNA
<213>芸香(Ruta graveolens)
<400>14
atgagtgcag cggcaacgtc gatgcaatcc cttaaattct ccaaccgtct ggtcccaccc 60
agtcgccgtc tgtctccggt tccgaacaat gtcacctgca ataacctccc caagtctgca 120
gctcccgtcc ggacagtcaa atgctgcgct tcttcctgga acagtaccat caacggcgcc 180
gccgccacga ccaacggtgc gtccgccgcc agtaacggcg catccacgac caccactaca 240
tatgttagtg atgcaaccag atttatcgac tcttc 275
<210>15
<211>162
<212>DNA
<213>玉蜀黍(Zea mays)
<400>15
atgatttcgc cgacgaatct cctcccggcg cggaagatca cccctgtctc aaatggcggc 60
gcagcgacgg cgagcccctc ttctccctcg gtggccgcac ggccacggcg actcccttca 120
ggcctccaat ctgtgactgg tagagggaag gtttccttgg ca 162
<210>16
<211>190
<212>DNA
<213>玉蜀黍(Zea mays)
<400>16
atgatttcgc cgacgaatct cctcccggcg cggaagatca cccctgtctc aaatggcggc 60
gcagcgacgg cgagcccctc ttctccctcg gtggccgcac ggccacggcg actcccttca 120
ggcctccaat ctgtgactgg tagagggaag gtttccttgg cagccatcac tctggatgat 180
taccttccaa 190
<210>17
<211>222
<212>DNA
<213>玉蜀黍(Zea mays)
<400>17
atgatttcgc cgacgaatct cctcccggcg cggaagatca cccctgtctc aaatggcggc 60
gcagcgacgg cgagcccctc ttctccctcg gtggccgcac ggccacggcg actcccttca 120
ggcctccaat ctgtgactgg tagagggaag gtttccttgg cagccatcac tctggatgat 180
taccttccaa tgcgaagcac tgaagtgaag aaccggacat ca 222
<210>18
<211>171
<212>DNA
<213>玉蜀黍(Zea mays)
<400>18
atgatttcgc cgacgaatct cctcccggcg cggaagatca cccctgtctc aaatggcggc 60
gcagcgacgg cgagcccctc ttctccctcg gtggccgcac ggccacggcg actcccttca 120
ggcctccaat ctgtgactgg tagagggaag gtttccttgg cagccatcac c 171
<210>19
<211>261
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>19
gcttcctcta tgctctcttc cgctactatg gttgcctctc cggctcaggc cactatggtc 60
gctcctttca acggacttaa gtcctccgct gccttcccag ccacccgcaa ggctaacaac 120
gacattactt ccatcacaag caacggcgga agagttaact gcatgcaggt gtggcctccg 180
attggaaaga agaagtttga gactctctct taccttcctg accttaccga ttccggtggt 240
cgcgtcaact gcatgcaggc c 261
<210>20
<211>76
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>20
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Cys
65 70 75
<210>21
<211>77
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>21
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Glu Lys
65 70 75
<210>22
<211>86
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>22
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Glu Lys Ala Ser Glu
65 70 75 80
Ile Val Leu Gln Pro Ile
85
<210>23
<211>81
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>23
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Glu Lys Ala Ser Glu
65 70 75 80
Ile
<210>24
<211>32
<212>PRT
<213>玉蜀黍(Zea mays)
<400>24
Met Ala Thr Ala Ser Leu Ala Leu Ser Leu Arg Leu Ala Pro Tyr Ser
1 5 10 15
His Pro Leu Ser Leu Arg Arg Arg Gly Ala Ala Gly Val Thr Cys Arg
20 25 30
<210>25
<211>52
<212>PRT
<213>玉蜀黍(Zea mays)
<400>25
Met Ala Thr Ala Ser Leu Ala Leu Ser Leu Arg Leu Ala Pro Tyr Ser
1 5 10 15
His Pro Leu Ser Leu Arg Arg Arg Gly Ala Ala Gly Val Thr Cys Arg
20 25 30
Ala Thr Thr Ala Thr Phe His Gln Leu Asp Ala Val Ala Val Arg Glu
35 40 45
Glu Glu Ser Arg
50
<210>26
<211>47
<212>PRT
<213>玉蜀黍(Zea mays)
<400>26
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser
35 40 45
<210>27
<211>63
<212>PRT
<213>玉蜀黍(Zea mays)
<400>27
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser Ala
35 40 45
Ala Val Thr Pro Met Val Thr Ile Ile Gln Asp Asp Gly Ala Glu
50 55 60
<210>28
<211>65
<212>PRT
<213>玉蜀黍(Zea mays)
<400>28
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser Ala
35 40 45
Ala Val Thr Pro Gln Ala Ser Pro Val Ile Ser Arg Ser Ala Ala Ala
50 55 60
Ala
65
<210>29
<211>75
<212>PRT
<213>稻(Oryza sativa)
<400>29
Ala Ala Leu Thr Thr Ser Gln Leu Ala Thr Ser Ala Thr Gly Phe Gly
1 5 10 15
Ile Ala Asp Arg Ser Ala Pro Ser Ser Leu Leu Arg His Gly Phe Gln
20 25 30
Gly Leu Lys Pro Arg Ser Pro Ala Gly Gly Asp Ala Thr Ser Leu Ser
35 40 45
Val Thr Thr Ser Ala Arg Ala Thr Pro Lys Gln Gln Arg Ser Val Gln
50 55 60
Arg Gly Ser Arg Arg Phe Pro Ser Val Val Val
65 70 75
<210>30
<211>80
<212>PRT
<213>稻(Oryza sativa)
<400>30
Ala Ala Leu Thr Thr Ser Gln Leu Ala Thr Ser Ala Thr Gly Phe Gly
1 5 10 15
Ile Ala Asp Arg Ser Ala Pro Ser Ser Leu Leu Arg His Gly Phe Gln
20 25 30
Gly Leu Lys Pro Arg Ser Pro Ala Gly Gly Asp Ala Thr Ser Leu Ser
35 40 45
Val Thr Thr Ser Ala Arg Ala Thr Pro Lys Gln Gln Arg Ser Val Gln
50 55 60
Arg Gly Ser Arg Arg Phe Pro Ser Val Val Val Tyr Ala Thr Gly Ala
65 70 75 80
<210>31
<211>96
<212>PRT
<213>稻(Oryza sativa)
<400>31
Ala Ala Leu Thr Thr Ser Gln Leu Ala Thr Ser Ala Thr Gly Phe Gly
1 5 10 15
Ile Ala Asp Arg Ser Ala Pro Ser Ser Leu Leu Arg His Gly Phe Gln
20 25 30
Gly Leu Lys Pro Arg Ser Pro Ala Gly Gly Asp Ala Thr Ser Leu Ser
35 40 45
Val Thr Thr Ser Ala Arg Ala Thr Pro Lys Gln Gln Arg Ser Val Gln
50 55 60
Arg Gly Ser Arg Arg Phe Pro Ser Val Val Val Tyr Ala Thr Gly Ala
65 70 75 80
Gly Met Asn Val Val Phe Val Gly Ala Glu Met Ala Pro Trp Ser Lys
85 90 95
<210>32
<211>71
<212>PRT
<213>芸香(Ruta graveolens)
<400>32
Met Ser Ala Ala Ala Thr Ser Met Gln Ser Leu Lys Phe Ser Asn Arg
1 5 10 15
Leu Val Pro Pro Ser Arg Arg Leu Ser Pro Val Pro Asn Asn Val Thr
20 25 30
Cys Asn Asn Leu Pro Lys Ser Ala Ala Pro Val Arg Thr Val Lys Cys
35 40 45
Cys Ala Ser Ser Trp Asn Ser Thr Ile Asn Gly Ala Ala Ala Thr Thr
50 55 60
Asn Gly Ala Ser Ala Ala Ser
65 70
<210>33
<211>91
<212>PRT
<213>芸香(Ruta graveolens)
<400>33
Met Ser Ala Ala Ala Thr Ser Met Gln Ser Leu Lys Phe Ser Asn Arg
1 5 10 15
Leu Val Pro Pro Ser Arg Arg Leu Ser Pro Val Pro Asn Asn Val Thr
20 25 30
Cys Asn Asn Leu Pro Lys Ser Ala Ala Pro Val Arg Thr Val Lys Cys
35 40 45
Cys Ala Ser Ser Trp Asn Ser Thr Ile Asn Gly Ala Ala Ala Thr Thr
50 55 60
Asn Gly Ala Ser Ala Ala Ser Asn Gly Ala Ser Thr Thr Thr Thr Thr
65 70 75 80
Tyr Val Ser Asp Ala Thr Arg Phe Ile Asp Ser
85 90
<210>34
<211>54
<212>PRT
<213>玉蜀黍(Zea mays)
<400>34
Met Ile Ser Pro Thr Asn Leu Leu Pro Ala Arg Lys Ile Thr Pro Val
1 5 10 15
Ser Asn Gly Gly Ala Ala Thr Ala Ser Pro Ser Ser Pro Ser Val Ala
20 25 30
Ala Arg Pro Arg Arg Leu Pro Ser Gly Leu Gln Ser Val Thr Gly Arg
35 40 45
Gly Lys Val Ser Leu Ala
50
<210>35
<211>63
<212>PRT
<213>玉蜀黍(Zea mays)
<400>35
Met Ile Ser Pro Thr Asn Leu Leu Pro Ala Arg Lys Ile Thr Pro Val
1 5 10 15
Ser Asn Gly Gly Ala Ala Thr Ala Ser Pro Ser Ser Pro Ser Val Ala
20 25 30
Ala Arg Pro Arg Arg Leu Pro Ser Gly Leu Gln Ser Val Thr Gly Arg
35 40 45
Gly Lys Val Ser Leu Ala Ala Ile Thr Leu Asp Asp Tyr Leu Pro
50 55 60
<210>36
<211>74
<212>PRT
<213>玉蜀黍(Zea mays)
<400>36
Met Ile Ser Pro Thr Asn Leu Leu Pro Ala Arg Lys Ile Thr Pro Val
1 5 10 15
Ser Asn Gly Gly Ala Ala Thr Ala Ser Pro Ser Ser Pro Ser Val Ala
20 25 30
Ala Arg Pro Arg Arg Leu Pro Ser Gly Leu Gln Ser Val Thr Gly Arg
35 40 45
Gly Lys Val Ser Leu Ala Ala Ile Thr Leu Asp Asp Tyr Leu Pro Met
50 55 60
Arg Ser Thr Glu Val Lys Asn Arg Thr Ser
65 70
<210>37
<211>57
<212>PRT
<213>玉蜀黍(Zea mays)
<400>37
Met Ile Ser Pro Thr Asn Leu Leu Pro Ala Arg Lys Ile Thr Pro Val
1 5 10 15
Ser Asn Gly Gly Ala Ala Thr Ala Ser Pro Ser Ser Pro Ser Val Ala
20 25 30
Ala Arg Pro Arg Arg Leu Pro Ser Gly Leu Gln Ser Val Thr Gly Arg
35 40 45
Gly Lys Val Ser Leu Ala Ala Ile Thr
50 55
<210>38
<211>87
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>38
Ala Ser Ser Met Leu Ser Ser Ala Thr Met Val Ala Ser Pro Ala Gln
1 5 10 15
Ala Thr Met Val Ala Pro Phe Asn Gly Leu Lys Ser Ser Ala Ala Phe
20 25 30
Pro Ala Thr Arg Lys Ala Asn Asn Asp Ile Thr Ser Ile Thr Ser Asn
35 40 45
Gly Gly Arg Val Asn Cys Met Gln Val Trp Pro Pro Ile Gly Lys Lys
50 55 60
Lys Phe Glu Thr Leu Ser Tyr Leu Pro Asp Leu Thr Asp Ser Gly Gly
65 70 75 80
Arg Val Asn Cys Met Gln Ala
85
<g10>39
<211>1134
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>39
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcgtgcat gggcaagggc 240
gaggaactgt tcactggcgt ggtcccaatc ctggtggaac tggatggtga tgtgaacggg 300
cacaagttct ccgtcagcgg agagggtgaa ggtgatgcca cctacggaaa gctcaccctg 360
aagttcatct gcactaccgg aaagctccct gttccgtggc caaccctcgt caccactttc 420
acctacggtg ttcagtgctt ctcccggtac ccagatcaca tgaagcagca tgacttcttc 480
aagagcgcca tgcccgaagg ctacgtgcaa gaaaggacta tctctttcaa ggatgacggg 540
aactacaaga cacgtgccga agtcaagttc gaaggtgata ccctggtgaa ccgcatcgag 600
ctgaaaggta agtttctgct tctacctttg atatatatat aataattatc attaattagt 660
agtaatataa tatttcaaat atttttttca aaataaaaga atgtagtata tagcaattgc 720
ttttctgtag tttataagtg tgtatatttt aatttataac ttttctaata tatgaccaaa 780
atttgttgat gtgcaggtat cgatttcaag gaagatggaa acatcctcgg acacaagctg 840
gagtacaact acaactccca caacgtatac atcacggccg acaagcagaa gaacggcatc 900
aaggctaact tcaagatcag gcacaacatc gaagatggaa gcgtgcaact ggcggaccac 960
taccagcaga acacgcccat cggcgatggc cctgtcctgc tgccggacaa ccattacctg 1020
tccacgcaat ctgccctctc caaggacccc aacgagaaga gggaccacat ggtcctgctg 1080
gagttcgtga cggctgctgg gatcacgcat ggcatggatg aactctacaa gtga 1134
<210>40
<211>1060
<212>DNA
<213>玉蜀黍(Zea mays)
<400>40
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc ggatccagga gcaaccatgg caagggcgag gaactgttca 180
ctggcgtggt cccaatcctg gtggaactgg atggtgatgt gaacgggcac aagttctccg 240
tcagcggaga gggtgaaggt gatgccacct acggaaagct caccctgaag ttcatctgca 300
ctaccggaaa gctccctgtt ccgtggccaa ccctcgtcac cactttcacc tacggtgttc 360
agtgcttctc ccggtaccca gatcacatga agcagcatga cttcttcaag agcgcatgcc 420
cgaaggctac gtgcaagaaa ggactatctc tttcaaggat gacgggaact acaagacacg 480
tgccgaagtc aagttcgaag gtgataccct ggtgaaccgc atcgagctga aaggtaagtt 540
tctgcttcta cctttgatat atatataata attatcatta attagtagta atataatatt 600
tcaaatattt ttttcaaaat aaaagaatgt agtatatagc aattgctttt ctgtagttta 660
taagtgtgta tattttaatt tataactttt ctaatatatg accaaaattt gttgatgtgc 720
aggtatcgat ttcaaggaag atggaaacat cctcggacac aagctggagt acaactacaa 780
ctcccacaac gtatacatca cggccgacaa gcagaagaac ggcatcaagg ctaacttcaa 840
gatcaggcac aacatcgaag atggaagcgt gcaactggcg gaccactacc agcagaacac 900
gcccatcggc gatggccctg tcctgctgcc ggacaaccat tacctgtcca cgcaatctgc 960
cctctccaag gaccccaacg agaagaggga ccacatggtc ctgctggagt tcgtgacggc 1020
tgctgggatc acgcatggca tggatgaact ctacaagtga 1060
<210>41
<211>606
<212>DNA
<213>稻(Oryza sativa)
<400>41
gcggctctca ccacgtccca gctcgccacc tcggccaccg gcttcggcat cgccgacagg 60
tcggcgccgt cgtcgctgct ccgccacggg ttccagggcc tcaagccccg cagccccgcc 120
ggcggcgacg cgacgtcgct cagcgtgacg accagcgcgc gcgcgacgcc caagcagcag 180
cggtcggtgc agcgtggcag ccggaggttc ccctccgtcg tcgtgtgatg ggcaagggcg 240
aggaactgtt cactggcgtg gtcccaatcc tggtggaact ggatggtgat gtgaacgggc 300
acaagttctc cgtcagcgga gagggtgaag gtgatgccac ctacggaaag ctcaccctga 360
agttcatctg cactaccgga aagctccctg ttccgtggcc aaccctcgtc accactttca 420
cctacggtgt tcagtgcttc tcccggtacc cagatcacat gaagcagcat gacttcttca 480
agagcgccat gcccgaaggc tacgtgcaag aaaggactat ctctttcaag gatgacggga 540
actacaagac acgtgccgaa gtcaagttcg aaggtgatac cctggtgaac cgcatcgagc 600
tgaaag 606
<210>42
<211>1140
<212>DNA
<213>芸香(Ruta graveolens)
<400>42
atgagtgcag cggcaacgtc gatgcaatcc cttaaattct ccaaccgtct ggtcccaccc 60
agtcgccgtc tgtctccggt tccgaacaat gtcacctgca ataacctccc caagtctgca 120
gctcccgtcc ggacagtcaa atgctgcgct tcttcctgga acagtaccat caacggcgcg 180
gccgccacga ccaacggtgc gtccgccgcc agttccgcgg atccaggagc aaccatgggc 240
aagggcgagg aactgttcac tggcgtggtc ccaatcctgg tggaactgga tggtgatgtg 300
aacgggcaca agttctccgt cagcggagag ggtgaaggtg atgccaccta cggaaagctc 360
accctgaagt tcatctgcac taccggaaag ctccctgttc cgtggccaac cctcgtcacc 420
actttcacct acggtgttca gtgcttctcc cggtacccag atcacatgaa gcagcatgac 480
ttcttcaaga gcgccatgcc cgaaggctac gtgcaagaaa ggactatctc tttcaaggat 540
gacgggaact acaagacacg tgccgaagtc aagttcgaag gtgataccct ggtgaaccgc 600
atcgagctga aaggtaagtt tctgcttcta cctttgatat atatataata attatcatta 660
attagtagta atataatatt tcaaatattt ttttcaaaat aaaagaatgt agtatatagc 720
aattgctttt ctgtagttta taagtgtgta tattttaatt tataactttt ctaatatatg 780
accaaaattt gttgatgtgc aggtatcgat ttcaaggaag atggaaacat cctcggacac 840
aagctggagt acaactacaa ctcccacaac gtatacatca cggccgacaa gcagaagaac 900
ggcatcaagg ctaacttcaa gatcaggcac aacatcgaag atggaagcgt gcaactggcg 960
gaccactacc agcagaacac gcccatcggc gatggccctg tcctgctgcc ggacaaccat 1020
tacctgtcca cgcaatctgc cctctccaag gaccccaacg agaagaggga ccacatggtc 1080
ctgctggagt tcgtgacggc tgctgggatc acgcatggca tggatgaact ctacaagtga 1140
<210>43
<211>1083
<212>DNA
<213>玉蜀黍(Zea mays)
<400>43
atgatttcgc cgacgaatct cctcccggcg cggaagatca cccctgtctc aaatggcggc 60
gcagcgacgg cgagcccctc ttctccctcg gtggccgcac ggccacggcg actcccttca 120
ggcctccaat ctgtgactgg tagagggaag gtttccttgg cggatccagg agcaaccatg 180
ggcaagggcg aggaactgtt cactggcgtg gtcccaatcc tggtggaact ggatggtgat 240
gtgaacgggc acaagttctc cgtcagcgga gagggtgaag gtgatgccac ctacggaaag 300
ctcaccctga agttcatctg cactaccgga aagctccctg ttccgtggcc aaccctcgtc 360
accactttca cctacggtgt tcagtgcttc tcccggtacc cagatcacat gaagcagcat 420
gacttcttca agagcgccat gcccgaaggc tacgtgcaag aaaggactat ctctttcaag 480
gatgacggga actacaagac acgtgccgaa gtcaagttcg aaggtgatac cctggtgaac 540
cgcatcgagc tgaaaggtaa gtttctgctt ctacctttga tatatatata ataattatca 600
ttaattagta gtaatataat atttcaaata tttttttcaa aataaaagaa tgtagtatat 660
agcaattgct tttctgtagt ttataagtgt gtatatttta atttataact tttctaatat 720
atgaccaaaa tttgttgatg tgcaggtatc gatttcaagg aagatggaaa catcctcgga 780
cacaagctgg agtacaacta caactcccac aacgtataca tcacggccga caagcagaag 840
aacggcatca aggctaactt caagatcagg cacaacatcg aagatggaag cgtgcaactg 900
gcggaccact accagcagaa cacgcccatc ggcgatggcc ctgtcctgct gccggacaac 960
cattacctgt ccacgcaatc tgccctctcc aaggacccca acgagaagag ggaccacatg 1020
gtcctgctgg agttcgtgac ggctgctggg atcacgcatg gcatggatga actctacaag 1080
tga 1083
<210>44
<211>1098
<212>DNA
<213>玉蜀黍(Zea mays)
<400>44
atgatttcgc cgacgaatct cctcccggcg cggaagatca cccctgtctc aaatggcggc 60
gcagcgacgg cgagcccctc ttctccctcg gtggccgcac ggccacggcg actcccttca 120
ggcctccaat ctgtgactgg tagagggaag gtttccttgg cagccatcac tctggaggat 180
ccaggagcaa ccatgggcaa gggcgaggaa ctgttcactg gcgtggtccc aatcctggtg 240
gaactggatg gtgatgtgaa cgggcacaag ttctccgtca gcggagaggg tgaaggtgat 300
gccacctacg gaaagctcac cctgaagttc atctgcacta ccggaaagct ccctgttccg 360
tggccaaccc tcgtcaccac tttcacctac ggtgttcagt gcttctcccg gtacccagat 420
cacatgaagc agcatgactt cttcaagagc gccatgcccg aaggctacgt gcaagaaagg 480
actatctctt tcaaggatga cgggaactac aagacacgtg ccgaagtcaa gttcgaaggt 540
gataccctgg tgaaccgcat cgagctgaaa ggtaagtttc tgcttctacc tttgatatat 600
atataataat tatcattaat tagtagtaat ataatatttc aaatattttt ttcaaaataa 660
aagaatgtag tatatagcaa ttgcttttct gtagtttata agtgtgtata ttttaattta 720
taacttttct aatatatgac caaaatttgt tgatgtgcag gtatcgattt caaggaagat 780
ggaaacatcc tcggacacaa gctggagtac aactacaact cccacaacgt atacatcacg 840
gccgacaagc agaagaacgg catcaaggct aacttcaaga tcaggcacaa catcgaagat 900
ggaagcgtgc aactggcgga ccactaccag cagaacacgc ccatcggcga tggccctgtc 960
ctgctgccgg acaaccatta cctgtccacg caatctgccc tctccaagga ccccaacgag 1020
aagagggacc acatggtcct gctggagttc gtgacggctg ctgggatcac gcatggcatg 1080
gatgaactct acaagtga 1098
<210>45
<211>2420
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>45
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcggagaa aatggcagcg 240
gtaattctag aagacggcgc ggagagttat accacgaagg gtggcatcgt cgtcacccgc 300
aggcggcgtg aggcatccta cagcgacgcg atcgccggtt atgtcgaccg gctggacgaa 360
cgccgcggcg cggtcttttc ctcgaactac gaatatcccg gccgctatac ccgctgggac 420
actgcggtgg tcgacccgcc gcttgccatc tcctccttcg gtcgctcgct ctggatcgaa 480
gcctataacg aacgcggcga agtgctgctg gcgctgatcg ccgaggatct gaagtccgtt 540
gccgacatca cgctcggctc acttgccgcc cgccgcctcg acctcaccat caacgagccc 600
gatcgtgtct tcaccgagga agagcggtcg aagatgccga cggtctttac ggttcttcgc 660
gcggtgacga acctcttcca ctcggaggag gactcgaacc tcggcctcta tggcgccttc 720
ggctacgacc tcgccttcca gttcgatgcg atcgaactga agctttcgcg tccggacgac 780
cagcgcgaca tggttctctt tctgccggac gagatccttg tggtcgatca ctatgcggcc 840
aaggcctgga tcgaccgcta cgatttcgcc agggagaacc tttcgaccga gggcaaggca 900
gcggacattg ctcccgagcc gttccgcagc gtcgacagca tcccgccgca cggggatcac 960
cgcccgggcg aatatgccga gctcgtcgtc aaggcgaagg aaagcttccg tcgcggcgat 1020
cttttcgaag tggtgccggg gcagaaattc tacgagcgct gcgaaagccg cccgtccgag 1080
atttccaacc ggctgaaggc gatcaatccg tcgccctatt ccttcttcat caatctcggc 1140
aaccaggaat atctcgtcgg tgcttcgccg gagatgttcg tgcgcgtttc cggccggcgc 1200
atcgagacct gcccgatctc cggtacgatc aagcgcggcg acgatccgat cgccgacagc 1260
gagcagatcc tgaagctctt gaactcgaag aaggacgagt ccgagctcac catgtgctcg 1320
gacgtcgacc gcaacgacaa gagccgggtc tgcgtgccgg gctcggtcaa ggtgatcggc 1380
cggcgtcaga tcgagatgta ttcgcggctg atccacacgg tcgatcacat cgaggggcgc 1440
ctgcgcgacg atatggacgc cttcgacggg ttcctcagcc acgcctgggc ggtgaccgtt 1500
accggcgcgc caaagctctg ggccatgcgc ttcatcgaga gccacgagaa gagcccgcgt 1560
gcctggtatg gcggcgcgat cggcatggtc ggcttcaacg gcgacatgaa taccgggctg 1620
accttgcgta ccatccgcat caaggacggg atcgccgagg tgagggcggg tgcgacgctc 1680
ctctatgatt ccaatccgga agaagaagaa gccgaaaccg aactgaaggc ctctgccatg 1740
attgcagcca tccgcgacgc gaaatccgca aacagcgcca aatccgcgcg cgatgtcgcc 1800
gccgtcggcg ccggagtcag catcctgctc gtcgatcacg aggacagctt cgtccatacc 1860
ctcgcgaact acttccgcca gaccggcgcg tccgtcacca ccgtgcgcac gccggtggcc 1920
gaggaaatct tcgaccgggt caagccggac ctcgtcgtgc tttcgcccgg tcccggcacc 1980
ccgaaggact tcgactgcaa ggcgacgatc aagaaggcgc gggcgcggga cctgccgatc 2040
ttcggcgtct gcctggggct gcaggcgctc gcggaggcct atggcggcga ccttcgtcaa 2100
ctggcgatcc cgatgcatgg gaagccctcg cgcatccgcg tgctcgaacc cggcatcgtc 2160
ttctccggcc tcggcaagga ggtgacggtc gggcgctatc attcgatttt cgccgatccg 2220
tccaacctgc cgcgcgaatt cgtgatcacg gccgaaagcg aagatggtac gatcatgggc 2280
atcgaacaca gcaaggagcc ggtggcggcc gtgcagttcc atccggaatc gatcatgacg 2340
ctgggcggcg acgccggcat gcggatgatc gagaacgtgg ttgcccatct cgccaagcgg 2400
gcgaagacca aggcagcctg 2420
<210>46
<211>2447
<212>DNA
<213>鼠耳芥(Arabidopsis thaliana)
<400>46
atggcgcaag ttagcagaat ctgcaatggt gtgcagaacc catctcttat ctccaatctc 60
tcgaaatcca gtcaacgcaa atctccctta tcggtttctc tgaagacgca gcagcatcca 120
cgagcttatc cgatttcgtc gtcgtgggga ttgaagaaga gtgggatgac gttaattggc 180
tctgagcttc gtcctcttaa ggtcatgtct tctgtttcca cggcggagaa agcgtcggag 240
attgtacttc aacccattat ggcagcggta attctagaag acggcgcgga gagttatacc 300
acgaagggtg gcatcgtcgt cacccgcagg cggcgtgagg catcctacag cgacgcgatc 360
gccggttatg tcgaccggct ggacgaacgc cgcggcgcgg tcttttcctc gaactacgaa 420
tatcccggcc gctatacccg ctgggacact gcggtggtcg acccgccgct tgccatctcc 480
tccttcggtc gctcgctctg gatcgaagcc tataacgaac gcggcgaagt gctgctggcg 540
ctgatcgccg aggatctgaa gtccgttgcc gacatcacgc tcggctcact tgccgcccgc 600
cgcctcgacc tcaccatcaa cgagcccgat cgtgtcttca ccgaggaaga gcggtcgaag 660
atgccgacgg tctttacggt tcttcgcgcg gtgacgaacc tcttccactc ggaggaggac 720
tcgaacctcg gcctctatgg cgccttcggc tacgacctcg ccttccagtt cgatgcgatc 780
gaactgaagc tttcgcgtcc ggacgaccag cgcgacatgg ttctctttct gccggacgag 840
atccttgtgg tcgatcacta tgcggccaag gcctggatcg accgctacga tttcgccagg 900
gagaaccttt cgaccgaggg caaggcagcg gacattgctc ccgagccgtt ccgcagcgtc 960
gacagcatcc cgccgcacgg ggatcaccgc ccgggcgaat atgccgagct cgtcgtcaag 1020
gcgaaggaaa gcttccgtcg cggcgatctt ttcgaagtgg tgccggggca gaaattctac 1080
gagcgctgcg aaagccgccc gtccgagatt tccaaccggc tgaaggcgat caatccgtcg 1140
ccctattcct tcttcatcaa tctcggcaac caggaatatc tcgtcggtgc ttcgccggag 1200
atgttcgtgc gcgtttccgg ccggcgcatc gagacctgcc cgatctccgg tacgatcaag 1260
cgcggcgacg atccgatcgc cgacagcgag cagatcctga agctcttgaa ctcgaagaag 1320
gacgagtccg agctcaccat gtgctcggac gtcgaccgca acgacaagag ccgggtctgc 1380
gtgccgggct cggtcaaggt gatcggccgg cgtcagatcg agatgtattc gcggctgatc 1440
cacacggtcg atcacatcga ggggcgcctg cgcgacgata tggacgcctt cgacgggttc 1500
ctcagccacg cctgggcggt gaccgttacc ggcgcgccaa agctctgggc catgcgcttc 1560
atcgagagcc acgagaagag cccgcgtgcc tggtatggcg gcgcgatcgg catggtcggc 1620
ttcaacggcg acatgaatac cgggctgacc ttgcgtacca tccgcatcaa ggacgggatc 1680
gccgaggtga gggcgggtgc gacgctcctc tatgattcca atccggaaga agaagaagcc 1740
gaaaccgaac tgaaggcctc tgccatgatt gcagccatcc gcgacgcgaa atccgcaaac 1800
agcgccaaat ccgcgcgcga tgtcgccgcc gtcggcgccg gagtcagcat cctgctcgtc 1860
gatcacgagg acagcttcgt ccataccctc gcgaactact tccgccagac cggcgcgtcc 1920
gtcaccaccg tgcgcacgcc ggtggccgag gaaatcttcg accgggtcaa gccggacctc 1980
gtcgtgcttt cgcccggtcc cggcaccccg aaggacttcg actgcaaggc gacgatcaag 2040
aaggcgcggg cgcgggacct gccgatcttc ggcgtctgcc tggggctgca ggcgctcgcg 2100
gaggcctatg gcggcgacct tcgtcaactg gcgatcccga tgcatgggaa gccctcgcgc 2160
atccgcgtgc tcgaacccgg catcgtcttc tccggcctcg gcaaggaggt gacggtcggg 2220
cgctatcatt cgattttcgc cgatccgtcc aacctgccgc gcgaattcgt gatcacggcc 2280
gaaagcgaag atggtacgat catgggcatc gaacacagca aggagccggt ggcggccgtg 2340
cagttccatc cggaatcgat catgacgctg ggcggcgacg ccggcatgcg gatgatcgag 2400
aacgtggttg cccatctcgc caagcgggcg aagaccaagg cagcctg 2447
<210>47
<211>2330
<212>DNA
<213>玉蜀黍(Zea mays)
<400>47
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc tatggcagcg gtaattctag aagacggcgc ggagagttat 180
accacgaagg gtggcatcgt cgtcacccgc aggcggcgtg aggcatccta cagcgacgcg 240
atcgccggtt atgtcgaccg gctggacgaa cgccgcggcg cggtcttttc ctcgaactac 300
gaatatcccg gccgctatac ccgctgggac actgcggtgg tcgacccgcc gcttgccatc 360
tcctccttcg gtcgctcgct ctggatcgaa gcctataacg aacgcggcga agtgctgctg 420
gcgctgatcg ccgaggatct gaagtccgtt gccgacatca cgctcggctc acttgccgcc 480
cgccgcctcg acctcaccat caacgagccc gatcgtgtct tcaccgagga agagcggtcg 540
aagatgccga cggtctttac ggttcttcgc gcggtgacga acctcttcca ctcggaggag 600
gactcgaacc tcggcctcta tggcgccttc ggctacgacc tcgccttcca gttcgatgcg 660
atcgaactga agctttcgcg tccggacgac cagcgcgaca tggttctctt tctgccggac 720
gagatccttg tggtcgatca ctatgcggcc aaggcctgga tcgaccgcta cgatttcgcc 780
agggagaacc tttcgaccga gggcaaggca gcggacattg ctcccgagcc gttccgcagc 840
gtcgacagca tcccgccgca cggggatcac cgcccgggcg aatatgccga gctcgtcgtc 900
aaggcgaagg aaagcttccg tcgcggcgat cttttcgaag tggtgccggg gcagaaattc 960
tacgagcgct gcgaaagccg cccgtccgag atttccaacc ggctgaaggc gatcaatccg 1020
tcgccctatt ccttcttcat caatctcggc aaccaggaat atctcgtcgg tgcttcgccg 1080
gagatgttcg tgcgcgtttc cggccggcgc atcgagacct gcccgatctc cggtacgatc 1140
aagcgcggcg acgatccgat cgccgacagc gagcagatcc tgaagctctt gaactcgaag 1200
aaggacgagt ccgagctcac catgtgctcg gacgtcgacc gcaacgacaa gagccgggtc 1260
tgcgtgccgg gctcggtcaa ggtgatcggc cggcgtcaga tcgagatgta ttcgcggctg 1320
atccacacgg tcgatcacat cgaggggcgc ctgcgcgacg atatggacgc cttcgacggg 1380
ttcctcagcc acgcctgggc ggtgaccgtt accggcgcgc caaagctctg ggccatgcgc 1440
ttcatcgaga gccacgagaa gagcccgcgt gcctggtatg gcggcgcgat cggcatggtc 1500
ggcttcaacg gcgacatgaa taccgggctg accttgcgta ccatccgcat caaggacggg 1560
atcgccgagg tgagggcggg tgcgacgctc ctctatgatt ccaatccgga agaagaagaa 1620
gccgaaaccg aactgaaggc ctctgccatg attgcagcca tccgcgacgc gaaatccgca 1680
aacagcgcca aatccgcgcg cgatgtcgcc gccgtcggcg ccggagtcag catcctgctc 1740
gtcgatcacg aggacagctt cgtccatacc ctcgcgaact acttccgcca gaccggcgcg 1800
tccgtcacca ccgtgcgcac gccggtggcc gaggaaatct tcgaccgggt caagccggac 1860
ctcgtcgtgc tttcgcccgg tcccggcacc ccgaaggact tcgactgcaa ggcgacgatc 1920
aagaaggcgc gggcgcggga cctgccgatc ttcggcgtct gcctggggct gcaggcgctc 1980
gcggaggcct atggcggcga ccttcgtcaa ctggcgatcc cgatgcatgg gaagccctcg 2040
cgcatccgcg tgctcgaacc cggcatcgtc ttctccggcc tcggcaagga ggtgacggtc 2100
gggcgctatc attcgatttt cgccgatccg tccaacctgc cgcgcgaatt cgtgatcacg 2160
gccgaaagcg aagatggtac gatcatgggc atcgaacaca gcaaggagcc ggtggcggcc 2220
gtgcagttcc atccggaatc gatcatgacg ctgggcggcg acgccggcat gcggatgatc 2280
gagaacgtgg ttgcccatct cgccaagcgg gcgaagacca aggcagcctg 2330
<210>48
<211>2384
<212>DNA
<213>玉蜀黍(Zea mays)
<400>48
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc tgctgccgtg acgccgcagg cgagcccagt gattagcagg 180
agcgctgcgg cggcgatggc agcggtaatt ctagaagacg gcgcggagag ttataccacg 240
aagggtggca tcgtcgtcac ccgcaggcgg cgtgaggcat cctacagcga cgcgatcgcc 300
ggttatgtcg accggctgga cgaacgccgc ggcgcggtct tttcctcgaa ctacgaatat 360
cccggccgct atacccgctg ggacactgcg gtggtcgacc cgccgcttgc catctcctcc 420
ttcggtcgct cgctctggat cgaagcctat aacgaacgcg gcgaagtgct gctggcgctg 480
atcgccgagg atctgaagtc cgttgccgac atcacgctcg gctcacttgc cgcccgccgc 540
ctcgacctca ccatcaacga gcccgatcgt gtcttcaccg aggaagagcg gtcgaagatg 600
ccgacggtct ttacggttct tcgcgcggtg acgaacctct tccactcgga ggaggactcg 660
aacctcggcc tctatggcgc cttcggctac gacctcgcct tccagttcga tgcgatcgaa 720
ctgaagcttt cgcgtccgga cgaccagcgc gacatggttc tctttctgcc ggacgagatc 780
cttgtggtcg atcactatgc ggccaaggcc tggatcgacc gctacgattt cgccagggag 840
aacctttcga ccgagggcaa ggcagcggac attgctcccg agccgttccg cagcgtcgac 900
agcatcccgc cgcacgggga tcaccgcccg ggcgaatatg ccgagctcgt cgtcaaggcg 960
aaggaaagct tccgtcgcgg cgatcttttc gaagtggtgc cggggcagaa attctacgag 1020
cgctgcgaaa gccgcccgtc cgagatttcc aaccggctga aggcgatcaa tccgtcgccc 1080
tattccttct tcatcaatct cggcaaccag gaatatctcg tcggtgcttc gccggagatg 1140
ttcgtgcgcg tttccggccg gcgcatcgag acctgcccga tctccggtac gatcaagcgc 1200
ggcgacgatc cgatcgccga cagcgagcag atcctgaagc tcttgaactc gaagaaggac 1260
gagtccgagc tcaccatgtg ctcggacgtc gaccgcaacg acaagagccg ggtctgcgtg 1320
ccgggctcgg tcaaggtgat cggccggcgt cagatcgaga tgtattcgcg gctgatccac 1380
acggtcgatc acatcgaggg gcgcctgcgc gacgatatgg acgccttcga cgggttcctc 1440
agccacgcct gggcggtgac cgttaccggc gcgccaaagc tctgggccat gcgcttcatc 1500
gagagccacg agaagagccc gcgtgcctgg tatggcggcg cgatcggcat ggtcggcttc 1560
aacggcgaca tgaataccgg gctgaccttg cgtaccatcc gcatcaagga cgggatcgcc 1620
gaggtgaggg cgggtgcgac gctcctctat gattccaatc cggaagaaga agaagccgaa 1680
accgaactga aggcctctgc catgattgca gccatccgcg acgcgaaatc cgcaaacagc 1740
gccaaatccg cgcgcgatgt cgccgccgtc ggcgccggag tcagcatcct gctcgtcgat 1800
cacgaggaca gcttcgtcca taccctcgcg aactacttcc gccagaccgg cgcgtccgtc 1860
accaccgtgc gcacgccggt ggccgaggaa atcttcgacc gggtcaagcc ggacctcgtc 1920
gtgctttcgc ccggtcccgg caccccgaag gacttcgact gcaaggcgac gatcaagaag 1980
gcgcgggcgc gggacctgcc gatcttcggc gtctgcctgg ggctgcaggc gctcgcggag 2040
gcctatggcg gcgaccttcg tcaactggcg atcccgatgc atgggaagcc ctcgcgcatc 2100
cgcgtgctcg aacccggcat cgtcttctcc ggcctcggca aggaggtgac ggtcgggcgc 2160
tatcattcga ttttcgccga tccgtccaac ctgccgcgcg aattcgtgat cacggccgaa 2220
agcgaagatg gtacgatcat gggcatcgaa cacagcaagg agccggtggc ggccgtgcag 2280
ttccatccgg aatcgatcat gacgctgggc ggcgacgccg gcatgcggat gatcgagaac 2340
gtggttgccc atctcgccaa gcgggcgaag accaaggcag cctg 2384
<210>49
<211>2379
<212>DNA
<213>玉蜀黍(Zea mays)
<400>49
atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt cccggcggcg 60
cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag ccggagcgga 120
accagcgggg tgaaatgctc tgctgccgtg acgccgcagg cgagcccagt gattagcagg 180
agcgctgcgg cggccgtaac gatcattcag gatgacggag cggagaccta cgagacgaaa 240
ggcggcatcc aggtcagccg aaagcgccgg cccaccgatt atgccaacgc catcgataat 300
tacatcgaaa agcttgattc ccatcgcggc gcggtttttt cgtccaacta tgaatatccg 360
ggccgttaca cccgctggga tacggccatc gtcgatccgc cgctcggcat ttcctgtttt 420
ggccgcaaga tgtggatcga agcctataat ggccgcggcg aagtgctgct cgatttcatt 480
acggaaaagc tgaaggcgac acccgatctc accctcggcg cttcctcgac ccgccggctc 540
gatcttaccg tcaacgaacc ggaccgtgtc ttcaccgaag aagaacgctc gaaaatcccg 600
acggtcttca ccgctctcag agccatcgtc gacctcttct attcgagcgc ggattcggcc 660
atcggcctgt tcggtgcctt cggttacgat ctcgccttcc agttcgacgc gatcaagctt 720
tcgctggcgc gtccggaaga ccagcgtgac atggtgctgt ttctgcccga tgaaatcctc 780
gtcgttgatc actattccgc caaggcctgg atcgaccgtt acgatttcga gaaggacggc 840
atgacgacgg acggcaaatc ctccgacatt acccccgatc ccttcaagac caccgatacc 900
atcccgccca agggcgatca ccgtcccggc gaatattccg agcttgtggt gaaggccaag 960
gaaagcttcc gccgcggcga cctgttcgag gtcgttcccg gccagaaatt catggagcgt 1020
tgcgaaagca atccgtcggc gatttcccgc cgcctgaagg cgatcaaccc gtcgccctat 1080
tccttcttca tcaatctcgg cgatcaggaa tatctggtcg gcgcctcgcc ggaaatgttc 1140
gtgcgcgtct ccggccgtcg catcgagacc tgcccgatat caggcaccat caagcgcggc 1200
gacgatccga ttgccgacag cgagcagatt ttgaaactgc tcaactcgaa aaaggacgaa 1260
tccgaactga ccatgtgctc ggacgtggac cgcaacgaca agagccgcgt ctgcgagccg 1320
ggttcggtga aggtcattgg ccgccgccag atcgagatgt attcacgcct catccacacc 1380
gtcgatcaca tcgaaggccg cctgcgcgac gatatggacg cctttgacgg tttcctcagc 1440
cacgcctggg ccgtcaccgt caccggtgca ccaaagctgt gggccatgcg cttcatcgaa 1500
ggtcatgaaa agagcccgcg cgcctggtat ggcggtgcga tcggcatggt cggcttcaac 1560
ggcgacatga ataccggcct gacgctgcgc accatccgga tcaaggacgg tattgccgaa 1620
gtgcgcgccg gcgcgaccct gctcaatgat tccaacccgc aggaagaaga agccgaaacc 1680
gaactgaagg cctccgccat gatatcagcc attcgtgacg caaaaggcac caactctgcc 1740
gccaccaagc gtgatgccgc caaagtcggc accggcgtca agatcctgct cgtcgaccac 1800
gaagacagct tcgtgcacac gctggcgaat tatttccgcc agacgggcgc gacggtctcg 1860
accgtcagat caccggtcgc agccgacgtg ttcgatcgct tccagccgga cctcgttgtc 1920
ctgtcgcccg gacccggcag cccgacggat ttcgactgca aggcaacgat caaggccgcc 1980
cgcgcccgcg atctgccgat cttcggcgtt tgcctcggtc tgcaggcatt ggcagaagcc 2040
tatggcggcg agctgcgcca gcttgctgtg cccatgcacg gcaagccttc gcgcatccgc 2100
gtgctggaac ccggcctcgt cttctccggt ctcggcaagg aagtcacggt cggtcgttac 2160
cattcgatct tcgccgatcc cgccaccctg ccgcgtgatt tcatcatcac cgcagaaagc 2220
gaggacggca cgatcatggg catcgaacac gccaaggaac cggtggccgc cgttcagttc 2280
cacccggaat cgatcatgac gctcggacag gacgcgggca tgcggatgat cgagaatgtc 2340
gtggtgcatc tgacccgcaa ggcgaagacc aaggccgcg 2379
<210>50
<211>314
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>50
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Cys Met Gly Lys Gly
65 70 75 80
Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly
85 90 95
Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp
100 105 110
Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys
115 120 125
Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe Thr Tyr Gly Val
130 135 140
Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe
145 150 155 160
Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Ser Phe
165 170 175
Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly
180 185 190
Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu
195 200 205
Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His
210 215 220
Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn
225 230 235 240
Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp
245 250 255
His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro
260 265 270
Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn
275 280 285
Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly
290 295 300
Ile Thr His Gly Met Asp Glu Leu Tyr Lys
305 310
<210>51
<211>290
<212>PRT
<213>玉蜀黍(Zea mays)
<400>51
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser Asp
35 40 45
Pro Gly Ala Thr Met Gly Lys Gly Glu Glu Leu Phe Thr Gly Val Val
50 55 60
Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser
65 70 75 80
Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu
85 90 95
Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu
100 105 110
Val Thr Thr Phe Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp
115 120 125
His Met Lys Gln His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr
130 135 140
Val Gln Glu Arg Thr Ile Ser Phe Lys Asp Asp Gly Asn Tyr Lys Thr
145 150 155 160
Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu
165 170 175
Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys
180 185 190
Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys
195 200 205
Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu
210 215 220
Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile
225 230 235 240
Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln
245 250 255
Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu
260 265 270
Leu Glu Phe Val Thr Ala Ala Gly Ile Thr His Gly Met Asp Glu Leu
275 280 285
Tyr Lys
290
<210>52
<211>201
<212>PRT
<213>稻(Oryza sativa)
<400>52
Ala Ala Leu Thr Thr Ser Gln Leu Ala Thr Ser Ala Thr Gly Phe Gly
1 5 10 15
Ile Ala Asp Arg Ser Ala Pro Ser Ser Leu Leu Arg His Gly Phe Gln
20 25 30
Gly Leu Lys Pro Arg Ser Pro Ala Gly Gly Asp Ala Thr Ser Leu Ser
35 40 45
Val Thr Thr Ser Ala Arg Ala Thr Pro Lys Gln Gln Arg Ser Val Gln
50 55 60
Arg Gly Ser Arg Arg Phe Pro Ser Val Val Val Met Gly Lys Gly Glu
65 70 75 80
Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp
85 90 95
Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala
100 105 110
Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu
115 120 125
Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe Thr Tyr Gly Val Gln
130 135 140
Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys
145 150 155 160
Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Ser Phe Lys
165 170 175
Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp
180 185 190
Thr Leu Val Asn Arg Ile Glu Leu Lys
195 200
<210>53
<211>316
<212>PRT
<213>芸香(Ruta graveolens)
<400>53
Met Ser Ala Ala Ala Thr Ser Met Gln Ser Leu Lys Phe Ser Asn Arg
1 5 10 15
Leu Val Pro Pro Ser Arg Arg Leu Ser Pro Val Pro Asn Asn Val Thr
20 25 30
Cys Asn Asn Leu Pro Lys Ser Ala Ala Pro Val Arg Thr Val Lys Cys
35 40 45
Cys Ala Ser Ser Trp Asn Ser Thr Ile Asn Gly Ala Ala Ala Thr Thr
50 55 60
Asn Gly Ala Ser Ala Ala Ser Ser Ala Asp Pro Gly Ala Thr Met Gly
65 70 75 80
Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu
85 90 95
Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu
100 105 110
Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr
115 120 125
Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe Thr Tyr
130 135 140
Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp
145 150 155 160
Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile
165 170 175
Ser Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe
180 185 190
Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe
195 200 205
Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn
210 215 220
Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys
225 230 235 240
Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu
245 250 255
Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu
260 265 270
Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp
275 280 285
Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala
290 295 300
Ala Gly Ile Thr His Gly Met Asp Glu Leu Tyr Lys
305 310 315
<210>54
<211>297
<212>PRT
<213>玉蜀黍(Zea mays)
<400>54
Met Ile Ser Pro Thr Asn Leu Leu Pro Ala Arg Lys Ile Thr Pro Val
1 5 10 15
Ser Asn Gly Gly Ala Ala Thr Ala Ser Pro Ser Ser Pro Ser Val Ala
20 25 30
Ala Arg Pro Arg Arg Leu Pro Ser Gly Leu Gln Ser Val Thr Gly Arg
35 40 45
Gly Lys Val Ser Leu Ala Asp Pro Gly Ala Thr Met Gly Lys Gly Glu
50 55 60
Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp
65 70 75 80
Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala
85 90 95
Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu
100 105 110
Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe Thr Tyr Gly Val Gln
115 120 125
Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln His Asp Phe Phe Lys
130 135 140
Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Ser Phe Lys
145 150 155 160
Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp
165 170 175
Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp
180 185 190
Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn
195 200 205
Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe
210 215 220
Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His
225 230 235 240
Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp
245 250 255
Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu
260 265 270
Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile
275 280 285
Thr His Gly Met Asp Glu Leu Tyr Lys
290 295
<210>55
<211>302
<212>PRT
<213>玉蜀黍(Zea mays)
<400>55
Met Ile Ser Pro Thr Asn Leu Leu Pro Ala Arg Lys Ile Thr Pro Val
1 5 10 15
Ser Asn Gly Gly Ala Ala Thr Ala Ser Pro Ser Ser Pro Ser Val Ala
20 25 30
Ala Arg Pro Arg Arg Leu Pro Ser Gly Leu Gln Ser Val Thr Gly Arg
35 40 45
Gly Lys Val Ser Leu Ala Ala Ile Thr Leu Glu Asp Pro Gly Ala Thr
50 55 60
Met Gly Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val
65 70 75 80
Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu
85 90 95
Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys
100 105 110
Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe
115 120 125
Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln
130 135 140
His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg
145 150 155 160
Thr Ile Ser Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val
165 170 175
Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile
180 185 190
Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn
195 200 205
Tyr Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly
210 215 220
Ile Lys Ala Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser Val
225 230 235 240
Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro
245 250 255
Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser
260 265 270
Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val
275 280 285
Thr Ala Ala Gly Ile Thr His Gly Met Asp Glu Leu Tyr Lys
290 295 300
<210>56
<211>806
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>56
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Glu Lys Met Ala Ala
65 70 75 80
Val Ile Leu Glu Asp Gly Ala Glu Ser Tyr Thr Thr Lys Gly Gly Ile
85 90 95
Val Val Thr Arg Arg Arg Arg Glu Ala Ser Tyr Ser Asp Ala Ile Ala
100 105 110
Gly Tyr Val Asp Arg Leu Asp Glu Arg Arg Gly Ala Val Phe Ser Ser
115 120 125
Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr Ala Val Val
130 135 140
Asp Pro Pro Leu Ala Ile Ser Ser Phe Gly Arg Ser Leu Trp Ile Glu
145 150 155 160
Ala Tyr Asn Glu Arg Gly Glu Val Leu Leu Ala Leu Ile Ala Glu Asp
165 170 175
Leu Lys Ser Val Ala Asp Ile Thr Leu Gly Ser Leu Ala Ala Arg Arg
180 185 190
Leu Asp Leu Thr Ile Asn Glu Pro Asp Arg Val Phe Thr Glu Glu Glu
195 200 205
Arg Ser Lys Met Pro Thr Val Phe Thr Val Leu Arg Ala Val Thr Asn
210 215 220
Leu Phe His Ser Glu Glu Asp Ser Asn Leu Gly Leu Tyr Gly Ala Phe
225 230 235 240
Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Glu Leu Lys Leu Ser
245 250 255
Arg Pro Asp Asp Gln Arg Asp Met Val Leu Phe Leu Pro Asp Glu Ile
260 265 270
Leu Val Val Asp His Tyr Ala Ala Lys Ala Trp Ile Asp Arg Tyr Asp
275 280 285
Phe Ala Arg Glu Asn Leu Ser Thr Glu Gly Lys Ala Ala Asp Ile Ala
290 295 300
Pro Glu Pro Phe Arg Ser Val Asp Ser Ile Pro Pro His Gly Asp His
305 310 315 320
Arg Pro Gly Glu Tyr Ala Glu Leu Val Val Lys Ala Lys Glu Ser Phe
325 330 335
Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys Phe Tyr Glu
340 345 350
Arg Cys Glu Ser Arg Pro Ser Glu Ile Ser Asn Arg Leu Lys Ala Ile
355 360 365
Asn Pro Ser Pro Tyr Ser Phe PheIle Asn Leu Gly Asn Gln Glu Tyr
370 375 380
Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser Gly Arg Arg
385 390 395 400
Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly Asp Asp Pro
405 410 415
Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser Lys Lys Asp
420 425 430
Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn Asp Lys Ser
435 440 445
Arg Val Cys Val Pro Gly Ser Val Lys Val Ile Gly Arg Arg Gln Ile
450 455 460
Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile Glu Gly Arg
465 470 475 480
Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser His Ala Trp
485 490 495
Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met Arg Phe Ile
500 505 510
Glu Ser His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly Ala Ile Gly
515 520 525
Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr Leu Arg Thr
530 535 540
Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly Ala Thr Leu
545 550 555 560
Leu Tyr Asp Ser Asn Pro Glu Glu Glu Glu Ala Glu Thr Glu Leu Lys
565 570 575
Ala Ser Ala Met Ile Ala Ala Ile Arg Asp Ala Lys Ser Ala Asn Ser
580 585 590
Ala Lys Ser Ala Arg Asp Val Ala Ala Val Gly Ala Gly Val Ser Ile
595 600 605
Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu Ala Asn Tyr
610 615 620
Phe Arg Gln Thr Gly Ala Ser Val Thr Thr Val Arg Thr Pro Val Ala
625 630 635 640
Glu Glu Ile Phe Asp Arg Val Lys Pro Asp Leu Val Val Leu Ser Pro
645 650 655
Gly Pro Gly Thr Pro Lys Asp Phe Asp Cys Lys Ala Thr Ile Lys Lys
660 665 670
Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu Gly Leu Gln
675 680 685
Ala Leu Ala Glu Ala Tyr Gly Gly Asp Leu Arg Gln Leu Ala Ile Pro
690 695 700
Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro Gly Ile Val
705 710 715 720
Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr His Ser Ile
725 730 735
Phe Ala Asp Pro Ser Asn Leu Pro Arg Glu Phe Val Ile Thr Ala Glu
740 745 750
Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ser Lys Glu Pro Val
755 760 765
Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu Gly Gly Asp
770 775 780
Ala Gly Met Arg Met Ile Glu Asn Val Val Ala His Leu Ala Lys Arg
785 790 795 800
Ala Lys Thr Lys Ala Ala
805
<210>57
<211>815
<212>PRT
<213>鼠耳芥(Arabidopsis thaliana)
<400>57
Met Ala Gln Val Ser Arg Ile Cys Asn Gly Val Gln Asn Pro Ser Leu
1 5 10 15
Ile Ser Asn Leu Ser Lys Ser Ser Gln Arg Lys Ser Pro Leu Ser Val
20 25 30
Ser Leu Lys Thr Gln Gln His Pro Arg Ala Tyr Pro Ile Ser Ser Ser
35 40 45
Trp Gly Leu Lys Lys Ser Gly Met Thr Leu Ile Gly Ser Glu Leu Arg
50 55 60
Pro Leu Lys Val Met Ser Ser Val Ser Thr Ala Glu Lys Ala Ser Glu
65 70 75 80
Ile Val Leu Gln Pro Ile Met Ala Ala Val Ile Leu Glu Asp Gly Ala
85 90 95
Glu Ser Tyr Thr Thr Lys Gly Gly Ile Val Val Thr Arg Arg Arg Arg
100 105 110
Glu Ala Ser Tyr Ser Asp Ala Ile Ala Gly Tyr Val Asp Arg Leu Asp
115 120 125
Glu Arg Arg Gly Ala Val Phe Ser Ser Asn Tyr Glu Tyr Pro Gly Arg
130 135 140
Tyr Thr Arg Trp Asp Thr Ala Val Val Asp Pro Pro Leu Ala Ile Ser
145 150 155 160
Ser Phe Gly Arg Ser Leu Trp Ile Glu Ala Tyr Asn Glu Arg Gly Glu
165 170 175
Val Leu Leu Ala Leu Ile Ala Glu Asp Leu Lys Ser Val Ala Asp Ile
180 185 190
Thr Leu Gly Ser Leu Ala Ala Arg Arg Leu Asp Leu Thr Ile Asn Glu
195 200 205
Pro Asp Arg Val Phe Thr Glu Glu Glu Arg Ser Lys Met Pro Thr Val
210 215 220
Phe Thr Val Leu Arg Ala Val Thr Asn Leu Phe His Ser Glu Glu Asp
225 230 235 240
Ser Asn Leu Gly Leu Tyr Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln
245 250 255
Phe Asp Ala Ile Glu Leu Lys Leu Ser Arg Pro Asp Asp Gln Arg Asp
260 265 270
Met Val Leu Phe Leu Pro Asp Glu Ile Leu Val Val Asp His Tyr Ala
275 280 285
Ala Lys Ala Trp Ile Asp Arg Tyr Asp Phe Ala Arg Glu Asn Leu Ser
290 295 300
Thr Glu Gly Lys Ala Ala Asp Ile Ala Pro Glu Pro Phe Arg Ser Val
305 310 315 320
Asp Ser Ile Pro Pro His Gly Asp His Arg Pro Gly Glu Tyr Ala Glu
325 330 335
Leu Val Val Lys Ala Lys Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu
340 345 350
Val Val Pro Gly Gln Lys Phe Tyr Glu Arg Cys Glu Ser Arg Pro Ser
355 360 365
Glu Ile Ser Asn Arg Leu Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe
370 375 380
Phe Ile Asn Leu Gly Asn Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu
385 390 395 400
Met Phe Val Arg Val Ser Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser
405 410 415
Gly Thr Ile Lys Arg Gly Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile
420 425 430
Leu Lys Leu Leu Asn Ser Lys Lys Asp Glu Ser Glu Leu Thr Met Cys
435 440 445
Ser Asp Val Asp Arg Asn Asp Lys Ser Arg Val Cys Val Pro Gly Ser
450 455 460
Val Lys Val Ile Gly Arg Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile
465 470 475 480
His Thr Val Asp His Ile Glu Gly Arg Leu Arg Asp Asp Met Asp Ala
485 490 495
Phe Asp Gly Phe Leu Ser His Ala Trp Ala Val Thr Val Thr Gly Ala
500 505 510
Pro Lys Leu Trp Ala Met Arg Phe Ile Glu Ser His Glu Lys Ser Pro
515 520 525
Arg Ala Trp Tyr Gly Gly Ala Ile Gly Met Val Gly Phe Asn Gly Asp
530 535 540
Met Asn Thr Gly Leu Thr Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile
545 550 555 560
Ala Glu Val Arg Ala Gly Ala Thr Leu Leu Tyr Asp Ser Asn Pro Glu
565 570 575
Glu Glu Glu Ala Glu Thr Glu Leu Lys Ala Ser Ala Met Ile Ala Ala
580 585 590
Ile Arg Asp Ala Lys Ser Ala Asn Ser Ala Lys Ser Ala Arg Asp Val
595 600 605
Ala Ala Val Gly Ala Gly Val Ser Ile Leu Leu Val Asp His Glu Asp
610 615 620
Ser Phe Val His Thr Leu Ala Asn Tyr Phe Arg Gln Thr Gly Ala Ser
625 630 635 640
Val Thr Thr Val Arg Thr Pro Val Ala Glu Glu Ile Phe Asp Arg Val
645 650 655
Lys Pro Asp Leu Val Val Leu Ser Pro Gly Pro Gly Thr Pro Lys Asp
660 665 670
Phe Asp Cys Lys Ala Thr Ile Lys Lys Ala Arg Ala Arg Asp Leu Pro
675 680 685
Ile Phe Gly Val Cys Leu Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly
690 695 700
Gly Asp Leu Arg Gln Leu Ala Ile Pro Met His Gly Lys Pro Ser Arg
705 710 715 720
Ile Arg Val Leu Glu Pro Gly Ile Val Phe Ser Gly Leu Gly Lys Glu
725 730 735
Val Thr Val Gly Arg Tyr His Ser Ile Phe Ala Asp Pro Ser Asn Leu
740 745 750
Pro Arg Glu Phe Val Ile Thr Ala Glu Ser Glu Asp Gly Thr Ile Met
755 760 765
Gly Ile Glu His Ser Lys Glu Pro Val Ala Ala Val Gln Phe His Pro
770 775 780
Glu Ser Ile Met Thr Leu Gly Gly Asp Ala Gly Met Arg Met Ile Glu
785 790 795 800
Asn Val Val Ala His Leu Ala Lys Arg Ala Lys Thr Lys Ala Ala
805 810 815
<210>58
<211>776
<212>PRT
<213>玉蜀黍(Zea mays)
<400>58
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser Met
35 40 45
Ala Ala Val Ile Leu Glu Asp Gly Ala Glu Ser Tyr Thr Thr Lys Gly
50 55 60
Gly Ile Val Val Thr Arg Arg Arg Arg Glu Ala Ser Tyr Ser Asp Ala
65 70 75 80
Ile Ala Gly Tyr Val Asp Arg Leu Asp Glu Arg Arg Gly Ala Val Phe
85 90 95
Ser Ser Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr Ala
100 105 110
Val Val Asp Pro Pro Leu Ala Ile Ser Ser Phe Gly Arg Ser Leu Trp
115 120 125
Ile Glu Ala Tyr Asn Glu Arg Gly Glu Val Leu Leu Ala Leu Ile Ala
130 135 140
Glu Asp Leu Lys Ser Val Ala Asp Ile Thr Leu Gly Ser Leu Ala Ala
145 150 155 160
Arg Arg Leu Asp Leu Thr Ile Asn Glu Pro Asp Arg Val Phe Thr Glu
165 170 175
Glu Glu Arg Ser Lys Met Pro Thr Val Phe Thr Val Leu Arg Ala Val
180 185 190
Thr Asn Leu Phe His Ser Glu Glu Asp Ser Asn Leu Gly Leu Tyr Gly
195 200 205
Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Glu Leu Lys
210 215 220
Leu Ser Arg Pro Asp Asp Gln Arg Asp Met Val Leu Phe Leu Pro Asp
225 230 235 240
Glu Ile Leu Val Val Asp His Tyr Ala Ala Lys Ala Trp Ile Asp Arg
245 250 255
Tyr Asp Phe Ala Arg Glu Asn Leu Ser Thr Glu Gly Lys Ala Ala Asp
260 265 270
Ile Ala Pro Glu Pro Phe Arg Ser Val Asp Ser Ile Pro Pro His Gly
275 280 285
Asp His Arg Pro Gly Glu Tyr Ala Glu Leu Val Val Lys Ala Lys Glu
290 295 300
Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys Phe
305 310 315 320
Tyr Glu Arg Cys Glu Ser Arg Pro Ser Glu Ile Ser Asn Arg Leu Lys
325 330 335
Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly Asn Gln
340 345 350
Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser Gly
355 360 365
Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly Asp
370 375 380
Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser Lys
385 390 395 400
Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn Asp
405 410 415
Lys Ser Arg Val Cys Val Pro Gly Ser Val Lys Val Ile Gly Arg Arg
420 425 430
Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile Glu
435 440 445
Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser His
450 455 460
Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met Arg
465 470 475 480
Phe Ile Glu Ser His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly Ala
485 490 495
Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr Leu
500 505 510
Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly Ala
515 520 525
Thr Leu Leu Tyr Asp Ser Asn Pro Glu Glu Glu Glu Ala Glu Thr Glu
530 535 540
Leu Lys Ala Ser Ala Met Ile Ala Ala Ile Arg Asp Ala Lys Ser Ala
545 550 555 560
Asn Ser Ala Lys Ser Ala Arg Asp Val Ala Ala Val Gly Ala Gly Val
565 570 575
Ser Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu Ala
580 585 590
Asn Tyr Phe Arg Gln Thr Gly Ala Ser Val Thr Thr Val Arg Thr Pro
595 600 605
Val Ala Glu Glu Ile Phe Asp Arg Val Lys Pro Asp Leu Val Val Leu
610 615 620
Ser Pro Gly Pro Gly Thr Pro Lys Asp Phe Asp Cys Lys Ala Thr Ile
625 630 635 640
Lys Lys Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu Gly
645 650 655
Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Asp Leu Arg Gln Leu Ala
660 665 670
Ile Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro Gly
675 680 685
Ile Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr His
690 695 700
Ser Ile Phe Ala Asp Pro Ser Asn Leu Pro Arg Glu Phe Val Ile Thr
705 710 715 720
Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ser Lys Glu
725 730 735
Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu Gly
740 745 750
Gly Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Ala His Leu Ala
755 760 765
Lys Arg Ala Lys Thr Lys Ala Ala
770 775
<210>59
<211>794
<212>PRT
<213>玉蜀黍(Zea mays)
<400>59
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser Ala
35 40 45
Ala Val Thr Pro Gln Ala Ser Pro Val Ile Ser Arg Ser Ala Ala Ala
50 55 60
Ala Met Ala Ala Val Ile Leu Glu Asp Gly Ala Glu Ser Tyr Thr Thr
65 70 75 80
Lys Gly Gly Ile Val Val Thr Arg Arg Arg Arg Glu Ala Ser Tyr Ser
85 90 95
Asp Ala Ile Ala Gly Tyr Val Asp Arg Leu Asp Glu Arg Arg Gly Ala
100 105 110
Val Phe Ser Ser Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp
115 120 125
Thr Ala Val Val Asp Pro Pro Leu Ala Ile Ser Ser Phe Gly Arg Ser
130 135 140
Leu Trp Ile Glu Ala Tyr Asn Glu Arg Gly Glu Val Leu Leu Ala Leu
145 150 155 160
Ile Ala Glu Asp Leu Lys Ser Val Ala Asp Ile Thr Leu Gly Ser Leu
165 170 175
Ala Ala Arg Arg Leu Asp Leu Thr Ile Asn Glu Pro Asp Arg Val Phe
180 185 190
Thr Glu Glu Glu Arg Ser Lys Met Pro Thr Val Phe Thr Val Leu Arg
195 200 205
Ala Val Thr Asn Leu Phe His Ser Glu Glu Asp Ser Asn Leu Gly Leu
210 215 220
Tyr Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Glu
225 230 235 240
Leu Lys Leu Ser Arg Pro Asp Asp Gln Arg Asp Met Val Leu Phe Leu
245 250 255
Pro Asp Glu Ile Leu Val Val Asp His Tyr Ala Ala Lys Ala Trp Ile
260 265 270
Asp Arg Tyr Asp Phe Ala Arg Glu Asn Leu Ser Thr Glu Gly Lys Ala
275 280 285
Ala Asp Ile Ala Pro Glu Pro Phe Arg Ser Val Asp Ser Ile Pro Pro
290 295 300
His Gly Asp His Arg Pro Gly Glu Tyr Ala Glu Leu Val Val Lys Ala
305 310 315 320
Lys Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln
325 330 335
Lys Phe Tyr Glu Arg Cys Glu Ser Arg Pro Ser Glu Ile Ser Asn Arg
340 345 350
Leu Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly
355 360 365
Asn Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val
370 375 380
Ser Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg
385 390 395 400
Gly Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn
405 410 415
Ser Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg
420 425 430
Asn Asp Lys Ser Arg Val Cys Val Pro Gly Ser Val Lys Val Ile Gly
435 440 445
Arg Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His
450 455 460
Ile Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu
465 470 475 480
Ser His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala
485 490 495
Met Arg Phe Ile Glu Ser His Glu Lys Ser Pro Arg Ala Trp Tyr Gly
500 505 510
Gly Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu
515 520 525
Thr Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala
530 535 540
Gly Ala Thr Leu Leu Tyr Asp Ser Asn Pro Glu Glu Glu Glu Ala Glu
545 550 555 560
Thr Glu Leu Lys Ala Ser Ala Met Ile Ala Ala Ile Arg Asp Ala Lys
565 570 575
Ser Ala Asn Ser Ala Lys Ser Ala Arg Asp Val Ala Ala Val Gly Ala
580 585 590
Gly Val Ser Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr
595 600 605
Leu Ala Asn Tyr Phe Arg Gln Thr Gly Ala Ser Val Thr Thr Val Arg
610 615 620
Thr Pro Val Ala Glu Glu Ile Phe Asp Arg Val Lys Pro Asp Leu Val
625 630 635 640
Val Leu Ser Pro Gly Pro Gly Thr Pro Lys Asp Phe Asp Cys Lys Ala
645 650 655
Thr Ile Lys Lys Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys
660 665 670
Leu Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Asp Leu Arg Gln
675 680 685
Leu Ala Ile Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu
690 695 700
Pro Gly Ile Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg
705 710 715 720
Tyr His Ser Ile Phe Ala Asp Pro Ser Asn Leu Pro Arg Glu Phe Val
725 730 735
Ile Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ser
740 745 750
Lys Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr
755 760 765
Leu Gly Gly Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Ala His
770 775 780
Leu Ala Lys Arg Ala Lys Thr Lys Ala Ala
785 790
<210>60
<211>793
<212>PRT
<213>玉蜀黍(Zea mays)
<400>60
Met Glu Ser Leu Ala Ala Thr Ser Val Phe Ala Pro Ser Arg Val Ala
1 5 10 15
Val Pro Ala Ala Arg Ala Leu Val Arg Ala Gly Thr Val Val Pro Thr
20 25 30
Arg Arg Thr Ser Ser Arg Ser Gly Thr Ser Gly Val Lys Cys Ser Ala
35 40 45
Ala Val Thr Pro Gln Ala Ser Pro Val Ile Ser Arg Ser Ala Ala Ala
50 55 60
Ala Val Thr Ile Ile Gln Asp Asp Gly Ala Glu Thr Tyr Glu Thr Lys
65 70 75 80
Gly Gly Ile Gln Val Ser Arg Lys Arg Arg Pro Thr Asp Tyr Ala Asn
85 90 95
Ala Ile Asp Asn Tyr Ile Glu Lys Leu Asp Ser His Arg Gly Ala Val
100 105 110
Phe Ser Ser Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
115 120 125
Ala Ile Val Asp Pro Pro Leu Gly Ile Ser Cys Phe Gly Arg Lys Met
130 135 140
Trp Ile Glu Ala Tyr Asn Gly Arg Gly Glu Val Leu Leu Asp Phe Ile
145 150 155 160
Thr Glu Lys Leu Lys Ala Thr Pro Asp Leu Thr Leu Gly Ala Ser Ser
165 170 175
Thr Arg Arg Leu Asp Leu Thr Val Asn Glu Pro Asp Arg Val Phe Thr
180 185 190
Glu Glu Glu Arg Ser Lys Ile Pro Thr Val Phe Thr Ala Leu Arg Ala
195 200 205
Ile Val Asp Leu Phe Tyr Ser Ser Ala Asp Ser Ala Ile Gly Leu Phe
210 215 220
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Lys Leu
225 230 235 240
Ser Leu Ala Arg Pro Glu Asp Gln Arg Asp Met Val Leu Phe Leu Pro
245 250 255
Asp Glu Ile Leu Val Val Asp His Tyr Ser Ala Lys Ala Trp Ile Asp
260 265 270
Arg Tyr Asp Phe Glu Lys Asp Gly Met Thr Thr Asp Gly Lys Ser Ser
275 280 285
Asp Ile Thr Pro Asp Pro Phe Lys Thr Thr Asp Thr Ile Pro Pro Lys
290 295 300
Gly Asp His Arg Pro Gly Glu Tyr Ser Glu Leu Val Val Lys Ala Lys
305 310 315 320
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
325 330 335
Phe Met Glu Arg Cys Glu Ser Asn Pro Ser Ala Ile Ser Arg Arg Leu
340 345 350
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly Asp
355 360 365
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
370 375 380
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
385 390 395 400
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
405 410 415
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
420 425 430
Asp Lys Ser Arg Val Cys Glu Pro Gly Ser Val Lys Val Ile Gly Arg
435 440 445
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
450 455 460
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
465 470 475 480
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
485 490 495
Arg Phe Ile Glu Gly His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
500 505 510
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
515 520 525
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
530 535 540
Ala Thr Leu Leu Asn Asp Ser Asn Pro Gln Glu Glu Glu Ala Glu Thr
545 550 555 560
Glu Leu Lys Ala Ser Ala Met Ile Ser Ala Ile Arg Asp Ala Lys Gly
565 570 575
Thr Asn Ser Ala Ala Thr Lys Arg Asp Ala Ala Lys Val Gly Thr Gly
580 585 590
Val Lys Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
595 600 605
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Thr Val Ser Thr Val Arg Ser
610 615 620
Pro Val Ala Ala Asp Val Phe Asp Arg Phe Gln Pro Asp Leu Val Val
625 630 635 640
Leu Ser Pro Gly Pro Gly Ser Pro Thr Asp Phe Asp Cys Lys Ala Thr
645 650 655
Ile Lys Ala Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
660 665 670
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Glu Leu Arg Gln Leu
675 680 685
Ala Val Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
690 695 700
Gly Leu Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
705 710 715 720
His Ser Ile Phe Ala Asp Pro Ala Thr Leu Pro Arg Asp Phe Ile Ile
725 730 735
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ala Lys
740 745 750
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
755 760 765
Gly Gln Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Val His Leu
770 775 780
Thr Arg Lys Ala Lys Thr Lys Ala Ala
785 790
<210>61
<211>31
<212>DNA
<213>人工序列
<220>
<223>引物
<400>61
tattggatcc agaacaatgg cttcctctat g 31
<210>62
<211>27
<212>DNA
<213>人工序列
<220>
<223>引物
<400>62
tattggatcc atggcctgca tgcagtt 27
<210>63
<211>27
<212>DNA
<213>人工序列
<220>
<223>引物
<400>63
tattggatcc gcggccttgg tcttcgc 27
<210>64
<211>38
<212>DNA
<213>人工序列
<220>
<223>引物
<400>64
tattggatcc agaacaatgg taacgatcat tcaggatg 38
<210>65
<211>36
<212>DNA
<213>人工序列
<220>
<223>引物
<400>65
tattggatcc agaacaatgg aatccctagc cgccac 36
<210>66
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>66
tattggatcc tctttgtcta caaaagctga ctc 33
<210>67
<211>37
<212>DNA
<213>人工序列
<220>
<223>引物
<400>67
tattggatcc agaacaatgg ctgccgtgac gccgcag 37
<210>68
<211>30
<212>DNA
<213>人工序列
<220>
<223>引物
<400>68
tattggatcc agaaccatgg cggctctcac 30
<210>69
<211>28
<212>DNA
<213>人工序列
<220>
<223>引物
<400>69
tattggatcc gcaccgtgaa gcatgcac 28
<210>70
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物
<400>70
tattggatcc agaaccatga gtgcagcggc aacg 34
<210>71
<211>29
<212>DNA
<213>人工序列
<220>
<223>引物
<400>71
tattggattc cgcggaactg gcggcggac 29
<210>72
<211>38
<212>DNA
<213>人工序列
<220>
<223>引物
<400>72
tttctagagg atccagaacc atgagtgcag cggcaacg 38
<210>73
<211>27
<212>DNA
<213>人工序列
<220>
<223>引物
<400>73
ttaacgtacg tctccgctcc gtcatcc 27
<210>74
<211>22
<212>DNA
<213>人工序列
<220>
<223>引物
<400>74
ctagtctaga atgatttcgc cg 22
<210>75
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>75
cgtctcgtac gtctccgctc cgtc 24
<210>76
<211>27
<212>DNA
<213>人工序列
<220>
<223>引物
<400>76
tgctctagaa tggcgcaagt tagcaga 27
<210>77
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物
<400>77
tcttctagaa ttaccgctgc catgcacgcc gt 32
<210>78
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物
<400>78
tcttctagaa ttaccgctgc cattttctcc gccgt 35
<210>79
<211>38
<212>DNA
<213>人工序列
<220>
<223>引物
<400>79
tcttctagaa ttaccgctgc cataatgggt tgaagtac 38
<210>80
<211>27
<212>DNA
<213>人工序列
<220>
<223>引物
<400>80
tgctctagaa tggaatccct agccgcc 27
<210>81
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物
<400>81
tcttctagaa ttaccgctgc catagagcat ttcac 35
<210>82
<211>21
<212>DNA
<213>人工序列
<220>
<223>引物
<400>82
tgctctagaa tggccaccgc c 21
<210>83
<211>38
<212>DNA
<213>人工序列
<220>
<223>引物
<400>83
tcttctagaa ttaccgctgc catcgccgcc gcagcgct 38
<210>84
<211>66
<212>DNA
<213>人工序列
<220>
<223>引物
<400>84
tattggatcc agaaccatgg taacgatcat tcaggatgac ggagcggaga cgtacgagac 60
gaaagg 66
<210>85
<211>41
<212>DNA
<213>人工序列
<220>
<223>引物
<400>85
tttccatggc gcaagttagc agaatctgca atggtgtgca g 41
<210>86
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>86
aacccatctc ttatctccaa tctctcgaaa tccagtcaac 40
<210>87
<211>39
<212>DNA
<213>人工序列
<220>
<223>引物
<400>87
gcaaatctcc cttatcggtt tctctgaaga cgcagcagc 39
<210>88
<211>41
<212>DNA
<213>人工序列
<220>
<223>引物
<400>88
atccacgagc ttatccgatt tcgtcgtcgt ggggattgaa g 41
<210>89
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>89
aagagtggga tgacgttaat tggctctgag cttcgtcctc 40
<210>90
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>90
ttaaggtcat gtcttctgtt tccacggcgt gcatggtaac 40
<210>91
<211>36
<212>DNA
<213>人工序列
<220>
<223>引物
<400>91
gatcattcag gatgacggag cggagacgta cgagac 36
<210>92
<211>19
<212>DNA
<213>人工序列
<220>
<223>引物
<400>92
gtctcgtacg tctccgctc 19
<210>93
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>93
cgtcatcctg aatgatcgtt accatgcacg ccgtggaaac 40
<210>94
<211>43
<212>DNA
<213>人工序列
<220>
<223>引物
<400>94
agaagacatg accttaagag gacgaagctc agagccaatt aac 43
<210>95
<211>39
<212>DNA
<213>人工序列
<220>
<223>引物
<400>95
gtcatcccac tcttcttcaa tccccacgac gacgaaatc 39
<210>96
<211>38
<212>DNA
<213>人工序列
<220>
<223>引物
<400>96
ggataagctc gtggatgctg ctgcgtcttc agagaaac 38
<210>97
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>97
cgataaggga gatttgcgtt gactggattt cgagagattg 40
<210>98
<211>39
<212>DNA
<213>人工序列
<220>
<223>引物
<400>98
gagataagag atgggttctg cacaccattg cagattctg 39
<210>99
<211>43
<212>DNA
<213>人工序列
<220>
<223>引物
<400>99
ttaaggtcat gtcttctgtt tccacggcgg agaaaatggt aac 43
<210>100
<211>43
<212>DNA
<213>人工序列
<220>
<223>引物
<400>100
cgtcatcctg aatgatcgtt accattttct ccgccgtgga aac 43
<210>101
<211>36
<212>DNA
<213>人工序列
<220>
<223>引物
<400>101
ttaaggtcat gtcttctgtt tccacggcgg agaaag 36
<210>102
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物
<400>102
cgtcggagat tgtacttcaa cccattatgg taac 34
<210>103
<211>37
<212>DNA
<213>人工序列
<220>
<223>引物
<400>103
cgtcatcctg aatgatcgta accataatgg gttgaag 37
<210>104
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>104
tacaatctcc gacgctttct ccgccgtgga aac 33
<210>105
<211>55
<212>DNA
<213>人工序列
<220>
<223>引物
<400>105
ttaaggtcat gtcttctgtt tccacggcgg agaaagcgtc ggagattatg gtaac 55
<210>106
<211>55
<212>DNA
<213>人工序列
<220>
<223>引物
<400>106
cgtcatcctg aatgatcgtt accataatct ccgacgcttt ctccgccgtg gaaac 55
<210>107
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>107
tctagaggat ccagaaccat ggccaccgcc agc 33
<210>108
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>108
ctgcgcctcg cgccgtactc gcacccgctg agc 33
<210>109
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物
<400>109
gccgcggcgc cgccggcgtc acctgccgcg cc 32
<210>110
<211>25
<212>DNA
<213>人工序列
<220>
<223>引物
<400>110
tggccatggt tctggatcct ctaga 25
<210>111
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物
<400>111
agtacggcgc gaggcgcagc gagagcgcga gg 32
<210>112
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物
<400>112
acgccggcgg cgccgcggcg gcggaggctc ag 32
<210>113
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>113
cgtcatcctg aatgatcgtt accatggcgc ggc 33
<210>114
<211>32
<212>DNA
<213>人工序列
<220>
<223>引物
<400>114
gccgcggcgc cgccggcgtc acctgccgcg cc 32
<210>115
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>115
accagcttga cgccgtcgcg gtgagggagg agc 33
<210>116
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>116
gcgacggcgt caagctggtg gaacgtggcg gtc 33
<210>117
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物
<400>117
cgtcatcctg aatgatcgtt accatcctgg actc 34
<210>118
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>118
ctagtctaga ggatccagaa ccatggaatc cctagccgcc 40
<210>119
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>119
acctccgtgt tcgcgccctc ccgcgtcgcc gtcccggcgg 40
<210>120
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>120
cgcgggccct ggttagggcg gggacggtgg taccaaccag 40
<210>121
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>121
gcggacgagc agccggagcg gaaccagcgg ggtgaaatgc 40
<210>122
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>122
tctatggtaa cgatcattca ggatgacgga gcggagacgt 40
<210>123
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>123
cgtctcgtac gtctccgctc cgtcatcctg aatgatcgtt 40
<210>124
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>124
accatagagc atttcacccc gctggttccg ctccggctgc 40
<210>125
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>125
tcgtccgcct ggttggtacc accgtccccg ccctaaccag 40
<210>126
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>126
ggcccgcgcc gccgggacgg cgacgcggga gggcgcgaac 40
<210>127
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>127
acggaggtgg cggctaggga ttccatggtt ctggatcctc 40
<210>128
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>128
tctgctgccg tgacgccgat ggtaacgatc attcaggatg 40
<210>129
<211>30
<212>DNA
<213>人工序列
<220>
<223>引物
<400>129
accatcggcg tcacggcagc agagcatttc 30
<210>130
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>130
ccccgctggt tccgctccgg ctgc 24
<210>131
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物
<400>131
tctgctgccg tgacgccgca ggcgagccca gtgat 35
<210>132
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物
<400>132
ggtaacgatc attcaggatg acggagcgga gacgt 35
<210>133
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物
<400>133
accatcactg ggctcgcctg cggcgtcacg gcagc 35
<210>134
<211>35
<212>DNA
<213>人工序列
<220>
<223>引物
<400>134
agagcatttc accccgctgg ttccgctccg gctgc 35
<210>135
<211>22
<212>DNA
<213>人工序列
<220>
<223>引物
<400>135
ctagtctaga atggaatccc ta 22
<210>136
<211>22
<212>DNA
<213>人工序列
<220>
<223>引物
<400>136
tcctggatcc ggcgtcacgg ca 22
<210>137
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>137
tctagaggat ccagaaccat ggcggctctc acc 33
<210>138
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>138
agctcgccac ctccgccacc ggcttcggca tcg 33
<210>139
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>139
gtcggcgccg tcgtcgctgc tccgccacgg gtt 33
<210>140
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>140
ctcaagccac gcagcccagc cggcggcgac gcc 33
<210>141
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>141
tcagcgtgac gaccagcgcg cgcgcgacgc cca 33
<210>142
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>142
cggtcggtgc agcgtcgcag ccggagcttc cca 33
<210>143
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物
<400>143
gtcgtgtgca tggtaacgat cattcaggat gacc 34
<210>144
<211>20
<212>DNA
<213>人工序列
<220>
<223>引物
<400>144
atggttctgg atcctctaga 20
<210>145
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>145
gtggccgagg tggcgagctg ggacgtggtg aga 33
<210>146
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>146
cagcgacgac ggcgccgacc tgtcggcgat gcc 33
<210>147
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>147
ctgggctgcg tggcttgagg ccctggaacc cgt 33
<210>148
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>148
gcgctggtcg tcacgctgag cgacgtcgcg tcg 33
<210>149
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>149
tgccacgctg caccgaccgc tgctgcttcg gcg 33
<210>150
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>150
atcgttacca tgcacacgac gacggatggg aac 33
<210>151
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>151
gtctcgtacg tctccgctcc gtcatcctga atg 33
<210>152
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>152
gtcgtgtacg ccaccggcgc cggcatggta acg 33
<210>153
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>153
atcattcagg atgacggagc ggagacgtac gag 33
<210>154
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>154
cgccggtggc gtacacgacg acggatggga acc 33
<210>155
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物
<400>155
gtctcgtacg tctccgctcc gtcatcctga atga 34
<210>156
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>156
gtcgtgtacg ccaccggcgc cggcatgaac gtc 33
<210>157
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>157
tcggcgccga gatggccccc tggagcaaga tgg 33
<210>158
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>158
atcattcagg atgacggagc ggagacgtac gag 33
<210>159
<211>33
<212>DNA
<213>人工序列
<220>
<223>引物
<400>159
gggccatctc ggcgccgacg aacacgacgt tca 33
<210>160
<211>34
<212>DNA
<213>人工序列
<220>
<223>引物
<400>160
gtctcgtacg tctccgctcc gtcatcctga atga 34
<210>161
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>161
ctagtctaga ggatccagaa ccatgatttc gccgacgaat 40
<210>162
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>162
ctcctcccgg cgcggaagat cacccctgtc tcaaatggcg 40
<210>163
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>163
gcgcagcgac ggcgagcccc tcttctccct cggtggccgc 40
<210>164
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>164
acggccacgg cgactccctt caggcctcca atctgtgact 40
<210>165
<211>45
<212>DNA
<213>人工序列
<220>
<223>引物
<400>165
ggtagaggga aggtttcctt ggcaatggta acgatcattc aggat 45
<210>166
<211>45
<212>DNA
<213>人工序列
<220>
<223>引物
<400>166
cgtctcgtac gtctccgctc cgtcatcctg aatgatcgtt accat 45
<210>167
<211>44
<212>DNA
<213>人工序列
<220>
<223>引物
<400>167
tgccaaggaa accttccctctaccagtcac agattggagg cctg 44
<210>168
<211>41
<212>DNA
<213>人工序列
<220>
<223>引物
<400>168
aagggagtcg ccgtggccgt gcggccaccg agggagaaga g 41
<210>169
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>169
gggctcgccg tcgctgcgcc gccatttgag acaggggtga 40
<210>170
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>170
tcttccgcgc cgggaggaga ttcgtcggcg aaatcatggt 40
<210>171
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>171
ggtagaggga aggtttcctt ggcagccatc actctggatg 40
<210>172
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>172
attaccttcc aatggtaacg atcattcagg atgacggagc 40
<210>173
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>173
accattggaa ggtaatcatc cagagtgatg gctgccaagg 40
<210>174
<211>36
<212>DNA
<213>人工序列
<220>
<223>引物
<400>174
aaaccttccc tctaccagtc acagattgga ggcctg 36
<210>175
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>175
attaccttcc aatgcgaagc actgaagtga agaaccggac 40
<210>176
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>176
atcaatggta acgatcattc aggatgacgg agcggagacg 40
<210>177
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>177
accattgatg tccggttctt cacttcagtg cttcgcattg 40
<210>178
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>178
gaaggtaatc atccagagtg atggctgcca aggaaacctt 40
<210>179
<211>29
<212>DNA
<213>人工序列
<220>
<223>引物
<400>179
ccctctacca gtcacagatt ggaggcctg 29
<210>180
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>180
ggtagaggga aggtttcctt ggcagccatc actctggata 40
<210>181
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>181
tggtaacgat cattcaggat gacggagcgg agacgtacga 40
<210>182
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>182
cgtctcgtac gtctccgctc cgtcatcctg aatgatcgtt 40
<210>183
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>183
accatatcca gagtgatggc tgccaaggaa accttccctc 40
<210>184
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>184
taccagtcac agattggagg cctg 24
<210>185
<211>22
<212>DNA
<213>人工序列
<220>
<223>引物
<400>185
ctagtctaga atgatttcgc cg 22
<210>186
<211>22
<212>DNA
<213>人工序列
<220>
<223>引物
<400>186
tcctggatcc tccagagtga tg 22
<210>187
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>187
ctagtctaga ggatccagaa ccatgatttc gccgacgaat 40
<210>188
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>188
ctcctcccgg cgcggaagat cacccctgtc tcaaatggcg 40
<210>189
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>189
gcgcagcgac ggcgagcccc tcttctccct cggtggccgc 40
<210>190
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>190
acggccacgg cgactccctt caggcctcca atctgtgact 40
<210191
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>191
gggctcgccg tcgctgcgcc gccatttgag acaggggtga 40
<210>192
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>192
tcttccgcgc cgggaggaga ttcgtcggcg aaatcatggt 40
<210>193
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>193
ggtagaggga aggtttcctt ggcagccatc actctggata 40
<210>194
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>194
tggtaacgat cattcaggat gacggagcgg agacgtacga 40
<210>195
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>195
cgtctcgtac gtctccgctc cgtcatcctg aatgatcgtt 40
<210>196
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>196
accatatcca gagtgatggc tgccaaggaa accttccctc 40
<210>197
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>197
taccagtcac agattggagg cctg 24
<210>198
<211>40
<212>DNA
<213>人工序列
<220>
<223>引物
<400>198
accattggaa ggtaatcatc cagagtgatg gctgccaagg 40
<210>199
<211>2190
<212>DNA
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>199
atggtaacga tcattcagga tgacggagcg gagacctacg agacgaaagg cggcatccag 60
gtcagccgaa agcgccggcc caccgattat gccaacgcca tcgataatta catcgaaaag 120
cttgattccc atcgcggcgc ggttttttcg tccaactatg aatatccggg ccgttacacc 180
cgctgggata cggccatcgt cgatccgccg ctcggcattt cctgttttgg ccgcaagatg 240
tggatcgaag cctataatgg ccgcggcgaa gtgctgctcg atttcattac ggaaaagctg 300
aaggcgacac ccgatctcac cctcggcgct tcctcgaccc gccggctcga tcttaccgtc 360
aacgaaccgg accgtgtctt caccgaagaa gaacgctcga aaatcccgac ggtcttcacc 420
gctctcagag ccatcgtcga cctcttctat tcgagcgcgg attcggccat cggcctgttc 480
ggtgccttcg gttacgatct cgccttccag ttcgacgcga tcaagctttc gctggcgcgt 540
ccggaagacc agcgtgacat ggtgctgttt ctgcccgatg aaatcctcgt cgttgatcac 600
tattccgcca aggcctggat cgaccgttac gatttcgaga aggacggcat gacgacggac 660
ggcaaatcct ccgacattac ccccgatccc ttcaagacca ccgataccat cccgcccaag 720
ggcgatcacc gtcccggcga atattccgag cttgtggtga aggccaagga aagcttccgc 780
cgcggcgacc tgttcgaggt cgttcccggc cagaaattca tggagcgttg cgaaagcaat 840
ccgtcggcga tttcccgccg cctgaaggcg atcaacccgt cgccctattc cttcttcatc 900
aatctcggcg atcaggaata tctggtcggc gcctcgccgg aaatgttcgt gcgcgtctcc 960
ggccgtcgca tcgagacctg cccgatatca ggcaccatca agcgcggcga cgatccgatt 1020
gccgacagcg agcagatttt gaaactgctc aactcgaaaa aggacgaatc cgaactgacc 1080
atgtgctcgg acgtggaccg caacgacaag agccgcgtct gcgagccggg ttcggtgaag 1140
gtcattggcc gccgccagat cgagatgtat tcacgcctca tccacaccgt cgatcacatc 1200
gaaggccgcc tgcgcgacga tatggacgcc tttgacggtt tcctcagcca cgcctgggcc 1260
gtcaccgtca ccggtgcacc aaagctgtgg gccatgcgct tcatcgaagg tcatgaaaag 1320
agcccgcgcg cctggtatgg cggtgcgatc ggcatggtcg gcttcaacgg cgacatgaat 1380
accggcctga cgctgcgcac catccggatc aaggacggta ttgccgaagt gcgcgccggc 1440
gcgaccctgc tcaatgattc caacccgcag gaagaagaag ccgaaaccga actgaaggcc 1500
tccgccatga tatcagccat tcgtgacgca aaaggcacca actctgccgc caccaagcgt 1560
gatgccgcca aagtcggcac cggcgtcaag atcctgctcg tcgaccacga agacagcttc 1620
gtgcacacgc tggcgaatta tttccgccag acgggcgcga cggtctcgac cgtcagatca 1680
ccggtcgcag ccgacgtgtt cgatcgcttc cagccggacc tcgttgtcct gtcgcccgga 1740
cccggcagcc cgacggattt cgactgcaag gcaacgatca aggccgcccg cgcccgcgat 1800
ctgccgatct tcggcgtttg cctcggtctg caggcattgg cagaagccta tggcggcgag 1860
ctgcgccagc ttgctgtgcc catgcacggc aagccttcgc gcatccgcgt gctggaaccc 1920
ggcctcgtct tctccggtct cggcaaggaa gtcacggtcg gtcgttacca ttcgatcttc 1980
gccgatcccg ccaccctgcc gcgtgatttc atcatcaccg cagaaagcga ggacggcacg 2040
atcatgggca tcgaacacgc caaggaaccg gtggccgccg ttcagttcca cccggaatcg 2100
atcatgacgc tcggacagga cgcgggcatg cggatgatcg agaatgtcgt ggtgcatctg 2160
acccgcaagg cgaagaccaa ggccgcgtga 2190
<210>200
<211>729
<212>PRT
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>200
Met Val Thr Ile Ile Gln Asp Asp Gly Ala Glu Thr Tyr Glu Thr Lys
1 5 10 15
Gly Gly Ile Gln Val Ser Arg Lys Arg Arg Pro Thr Asp Tyr Ala Asn
20 25 30
Ala Ile Asp Asn Tyr Ile Glu Lys Leu Asp Ser His Arg Gly Ala Val
35 40 45
Phe Ser Ser Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
50 55 60
Ala Ile Val Asp Pro Pro Leu Gly Ile Ser Cys Phe Gly Arg Lys Met
65 70 75 80
Trp Ile Glu Ala Tyr Asn Gly Arg Gly Glu Val Leu Leu Asp Phe Ile
85 90 95
Thr Glu Lys Leu Lys Ala Thr Pro Asp Leu Thr Leu Gly Ala Ser Ser
100 105 110
Thr Arg Arg Leu Asp Leu Thr Val Asn Glu Pro Asp Arg Val Phe Thr
115 120 125
Glu Glu Glu Arg Ser Lys Ile Pro Thr Val Phe Thr Ala Leu Arg Ala
130 135 140
Ile Val Asp Leu Phe Tyr Ser Ser Ala Asp Ser Ala Ile Gly Leu Phe
145 150 155 160
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Lys Leu
165 170 175
Ser Leu Ala Arg Pro Glu Asp Gln Arg Asp Met Val Leu Phe Leu Pro
180 185 190
Asp Glu Ile Leu Val Val Asp His Tyr Ser Ala Lys Ala Trp Ile Asp
195 200 205
Arg Tyr Asp Phe Glu Lys Asp Gly Met Thr Thr Asp Gly Lys Ser Ser
210 215 220
Asp Ile Thr Pro Asp Pro Phe Lys Thr Thr Asp Thr Ile Pro Pro Lys
225 230 235 240
Gly Asp His Arg Pro Gly Glu Tyr Ser Glu Leu Val Val Lys Ala Lys
245 250 255
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
260 265 270
Phe Met Glu Arg Cys Glu Ser Asn Pro Ser Ala Ile Ser Arg Arg Leu
275 280 285
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe PheIle Asn Leu Gly Asp
290 295 300
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
305 310 315 320
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
325 330 335
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
340 345 350
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
355 360 365
Asp Lys Ser Arg Val Cys Glu Pro Gly Ser Val Lys Val Ile Gly Arg
370 375 380
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
385 390 395 400
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
405 410 415
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
420 425 430
Arg Phe Ile Glu Gly His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
435 440 445
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
450 455 460
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
465 470 475 480
Ala Thr Leu Leu Asn Asp Ser Asn Pro Gln Glu Glu Glu Ala Glu Thr
485 490 495
Glu Leu Lys Ala Ser Ala Met Ile Ser Ala Ile Arg Asp Ala Lys Gly
500 505 510
Thr Asn Ser Ala Ala Thr Lys Arg Asp Ala Ala Lys Val Gly Thr Gly
515 520 525
Val Lys Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
530 535 540
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Thr Val Ser Thr Val Arg Ser
545 550 555 560
Pro Val Ala Ala Asp Val Phe Asp Arg Phe Gln Pro Asp Leu Val Val
565 570 575
Leu Ser Pro Gly Pro Gly Ser Pro Thr Asp Phe Asp Cys Lys Ala Thr
580 585 590
Ile Lys Ala Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
595 600 605
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Glu Leu Arg Gln Leu
610 6l5 620
Ala Val Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
625 630 635 640
Gly Leu Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
645 650 655
His Ser Ile Phe Ala Asp Pro Ala Thr Leu Pro Arg Asp Phe Ile Ile
660 665 670
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ala Lys
675 680 685
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
690 695 700
Gly Gln Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Val His Leu
705 710 715 720
Thr Arg Lys Ala Lys Thr Lys Ala Ala
725
<210>201
<211>2190
<212>DNA
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>201
atggtaacga tcattcagga tgacggagcg gagacctacg agacgaaagg cggcatccag 60
gtcagccgaa agcgccggcc caccgattat gccaacgcca tcgataatta catcgaaaag 120
cttgattccc atcgcggcgc ggttttttcg tccaactatg aatatccggg ccgttacacc 180
cgctgggata cggccatcgt cgatccgccg ctcggcattt cctgttttgg ccgcaagatg 240
tggatcgaag cctataatgg ccgcggcgaa gtgctgctcg atttcattac ggaaaagctg 300
aaggcgacac ccgatctcac cctcggcgct tcctcgaccc gccggctcga tcttaccgtc 360
aacgaaccgg accgtgtctt caccgaagaa gaacgctcga aaatcccgac ggtcttcacc 420
gctctcagag ccatcgtcga cctcttctat tcgagcgcgg attcggccat cggcctgttc 480
ggtgccttcg gttacgatct cgccttccag ttcgacgcga tcaagctttc gctggcgcgt 540
ccggaagacc agcgtgacat ggtgctgttt ctgcccgatg aaatcctcgt cgttgatcac 600
tattccgcca aggcctggat cgaccgttac gatttcgaga aggacggcat gacgacggac 660
ggcaaatcct ccgacattac ccccgatccc ttcaagacca ccgataccat cccgcccaag 720
ggcgatcacc gtcccggcga atattccgag cttgtggtga aggccaagga aagcttccgc 780
cgcggcgacc tgttcgaggt cgttcccggc cagaaattca tggagcgttg cgaaagcaat 840
ccgtcggcga tttcccgccg cctgaaggcg atcaacccgt cgccctattc ctggttcatc 900
aatctcggcg atcaggaata tctggtcggc gcctcgccgg aaatgttcgt gcgcgtctcc 960
ggccgtcgca tcgagacctg cccgatatca ggcaccatca agcgcggcga cgatccgatt 1020
gccgacagcg agcagatttt gaaactgctc aactcgaaaa aggacgaatc cgaactgacc 1080
atgtgctcgg acgtggaccg caacgacaag agccgcgtct gcgagccggg ttcggtgaag 1140
gtcattggcc gccgccagat cgagatgtat tcacgcctca tccacaccgt cgatcacatc 1200
gaaggccgcc tgcgcgacga tatggacgcc tttgacggtt tcctcagcca cgcctgggcc 1260
gtcaccgtca ccggtgcacc aaagctgtgg gccatgcgct tcatcgaagg tcatgaaaag 1320
agcccgcgcg cctggtatgg cggtgcgatc ggcatggtcg gcttcaacgg cgacatgaat 1380
accggcctga cgctgcgcac catccggatc aaggacggta ttgccgaagt gcgcgccggc 1440
gcgaccctgc tcaatgattc caacccgcag gaagaagaag ccgaaaccga actgaaggcc 1500
tccgccatga tatcagccat tcgtgacgca aaaggcacca actctgccgc caccaagcgt 1560
gatgccgcca aagtcggcac cggcgtcaag atcctgctcg tcgaccacga agacagcttc 1620
gtgcacacgc tggcgaatta tttccgccag acgggcgcga cggtctcgac cgtcagatca 1680
ccggtcgcag ccgacgtgtt cgatcgcttc cagccggacc tcgttgtcct gtcgcccgga 1740
cccggcagcc cgacggattt cgactgcaag gcaacgatca aggccgcccg cgcccgcgat 1800
ctgccgatct tcggcgtttg cctcggtctg caggcattgg cagaagccta tggcggcgag 1860
ctgcgccagc ttgctgtgcc catgcacggc aagccttcgc gcatccgcgt gctggaaccc 1920
ggcctcgtct tctccggtct cggcaaggaa gtcacggtcg gtcgttacca ttcgatcttc 1980
gccgatcccg ccaccctgcc gcgtgatttc atcatcaccg cagaaagcga ggacggcacg 2040
atcatgggca tcgaacacgc caaggaaccg gtggccgccg ttcagttcca cccggaatcg 2100
atcatgacgc tcggacagga cgcgggcatg cggatgatcg agaatgtcgt ggtgcatctg 2160
acccgcaagg cgaagaccaa ggccgcgtga 2190
<2l0>202
<211>729
<212>PRT
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>202
Met Val Thr Ile Ile Gln Asp Asp Gly Ala Glu Thr Tyr Glu Thr Lys
1 5 10 15
Gly Gly Ile Gln Val Ser Arg Lys Arg Arg Pro Thr Asp Tyr Ala Asn
20 25 30
Ala Ile Asp Asn Tyr Ile Glu Lys Leu Asp Ser His Arg Gly Ala Val
35 40 45
Phe Ser Ser Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
50 55 60
Ala Ile Val Asp Pro Pro Leu Gly Ile Ser Cys Phe Gly Arg Lys Met
65 70 75 80
Trp Ile Glu Ala Tyr Asn Gly Arg Gly Glu Val Leu Leu Asp Phe Ile
85 90 95
Thr Glu Lys Leu Lys Ala Thr Pro Asp Leu Thr Leu Gly Ala Ser Ser
100 105 110
Thr Arg Arg Leu Asp Leu Thr Val Asn Glu Pro Asp Arg Val Phe Thr
115 120 125
Glu Glu Glu Arg Ser Lys Ile Pro Thr Val Phe Thr Ala Leu Arg Ala
130 135 140
Ile Val Asp Leu Phe Tyr Ser Ser Ala Asp Ser Ala Ile Gly Leu Phe
145 150 155 160
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Lys Leu
165 170 175
Ser Leu Ala Arg Pro Glu Asp Gln Arg Asp Met Val Leu Phe Leu Pro
180 185 190
Asp Glu Ile Leu Val Val Asp His Tyr Ser Ala Lys Ala Trp Ile Asp
195 200 205
Arg Tyr Asp Phe Glu Lys Asp Gly Met Thr Thr Asp Gly Lys Ser Ser
210 215 220
Asp Ile Thr Pro Asp Pro Phe Lys Thr Thr Asp Thr Ile Pro Pro Lys
225 230 235 240
Gly Asp His Arg Pro Gly Glu Tyr Ser Glu Leu Val Val Lys Ala Lys
245 250 255
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
260 265 270
Phe Met Glu Arg Cys Glu Ser Asn Pro Ser Ala Ile Ser Arg Arg Leu
275 280 285
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Trp Phe Ile Asn Leu Gly Asp
290 295 300
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
305 310 315 320
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
325 330 335
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
340 345 350
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
355 360 365
Asp Lys Ser Arg Val Cys Glu Pro Gly Ser Val Lys Val Ile Gly Arg
370 375 380
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
385 390 395 400
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
405 410 415
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
420 425 430
Arg Phe Ile Glu Gly His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
435 440 445
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
450 455 460
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
465 470 475 480
Ala Thr Leu Leu Asn Asp Ser Asn Pro Gln Glu Glu Glu Ala Glu Thr
485 490 495
Glu Leu Lys Ala Ser Ala Met Ile Ser Ala Ile Arg Asp Ala Lys Gly
500 505 510
Thr Asn Ser Ala Ala Thr Lys Arg Asp Ala Ala Lys Val Gly Thr Gly
515 520 525
Val Lys Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
530 535 540
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Thr Val Ser Thr Val Arg Ser
545 550 555 560
Pro Val Ala Ala Asp Val Phe Asp Arg Phe Gln Pro Asp Leu Val Val
565 570 575
Leu Ser Pro Gly Pro Gly Ser Pro Thr Asp Phe Asp Cys Lys Ala Thr
580 585 590
Ile Lys Ala Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
595 600 605
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Glu Leu Arg Gln Leu
610 615 620
Ala Val Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
625 630 635 640
Gly Leu Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
645 650 655
His Ser Ile Phe Ala Asp Pro Ala Thr Leu Pro Arg Asp Phe Ile Ile
660 665 670
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ala Lys
675 680 685
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
690 695 700
Gly Gln Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Val His Leu
705 710 715 720
Thr Arg Lys Ala Lys Thr Lys Ala Ala
725
<210>203
<211>2190
<212>DNA
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>203
gtaacgatca ttcaggatga cggagcggag acctacgaga cgaaaggcgg catccaggtc 60
agccgaaagc gccggcccac cgattatgcc aacgccatcg ataattacat cgaaaagctt 120
gattcccatc gcggcgcggt tttttcgttc aactatgaat atccgggccg ttacacccgc 180
tgggatacgg ccatcgtcga tccgccgctc ggcatttcct gttttggccg caagatgtgg 240
atcgaagcct ataatggccg cggcgaagtg ctgctcgatt tcattacgga aaagctgaag 300
gcgacacccg atctcaccct cggcgcttcc tcgacccgcc ggctcgatct taccgtcaac 360
gaaccggacc gtgtcttcac cgaagaagaa cgctcgaaaa tcccgacggt cttcaccgct 420
ctcagagcca tcgtcgacct cttctattcg agcgcggatt cggccatcgg cctgttcggt 480
gccttcggtt acgatctcgc cttccagttc gacgcgatca agctttcgct ggcgcgtccg 540
gaagaccagc gtgacatggt gctgtttctg cccgatgaaa tcctcgtcgt tgatcactat 600
tccgccaagg cctggatcga ccgttacgat ttcgagaagg acggcatgac gacggacggc 660
aaatcctccg acattacccc cgatcccttc aagaccaccg ataccatccc gcccaagggc 720
gatcaccgtc ccggcgaata ttccgagctt gtggtgaagg ccaaggaaag cttccgccgc 780
ggcgacctgt tcgaggtcgt tcccggccag aaattcatgg agcgttgcga aagcaatccg 840
tcggcgattt cccgccgcct gaaggcgatc aacccgtcgc cctattcctt cttcatcaat 900
ctcggcgatc aggaatatct ggtcggcgcc tcgccggaaa tgttcgtgcg cgtctccggc 960
cgtcgcatcg agacctgccc gatatcaggc accatcaagc gcggcgacga tccgattgcc 1020
gacagcgagc agattttgaa actgctcaac tcgaaaaagg acgaatccga actgaccatg 1080
tgctcggacg tggaccgcaa cgacaagagc cgcgtctgcg agccgggttc ggtgaaggtc 1140
attggccgcc gccagatcga gatgtattca cgcctcatcc acaccgtcga tcacatcgaa 1200
ggccgcctgc gcgacgatat ggacgccttt gacggtttcc tcagccacgc ctgggccgtc 1260
accgtcaccg gtgcaccaaa gctgtgggcc atgcgcttca tcgaaggtca tgaaaagagc 1320
ccgcgcgcct ggtatggcgg tgcgatcggc atggtcggct tcaacggcga catgaatacc 1380
ggcctgacgc tgcgcaccat ccggatcaag gacggtattg ccgaagtgcg cgccggcgcg 1440
accctgctca atgattccaa cccgcaggaa gaagaagccg aaaccgaact gaaggcctcc 1500
gccatgatat cagccattcg tgacgcaaaa ggcaccaact ctgccgccac caagcgtgat 1560
gccgccaaag tcggcaccgg cgtcaagatc ctgctcgtcg accacgaaga cagcttcgtg 1620
cacacgctgg cgaattattt ccgccagacg ggcgcgacgg tctcgaccgt cagatcaccg 1680
gtcgcagccg acgtgttcga tcgcttccag ccggacctcg ttgtcctgtc gcccggaccc 1740
ggcagcccga cggatttcga ctgcaaggca acgatcaagg ccgcccgcgc ccgcgatctg 1800
ccgatcttcg gcgtttgcct cggtctgcag gcattggcag aagcctatgg cggcgagctg 1860
cgccagcttg ctgtgcccat gcacggcaag ccttcgcgca tccgcgtgct ggaacccggc 1920
ctcgtcttct ccggtctcgg caaggaagtc acggtcggtc gttaccattc gatcttcgcc 1980
gatcccgcca ccctgccgcg tgatttcatc atcaccgcag aaagcgagga cggcacgatc 2040
atgggcatcg aacacgccaa ggaaccggtg gccgccgttc agttccaccc ggaatcgatc 2100
atgacgctcg gacaggacgc gggcatgcgg atgatcgaga atgtcgtggt gcatctgacc 2160
cgcaaggcga agaccaaggc cgcgtgatgg 2190
<210>204
<211>728
<212>PRT
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>204
Val Thr Ile Ile Gln Asp Asp Gly Ala Glu Thr Tyr Glu Thr Lys Gly
1 5 10 15
Gly Ile Gln Val Ser Arg Lys Arg Arg Pro Thr Asp Tyr Ala Asn Ala
20 25 30
Ile Asp Asn Tyr Ile Glu Lys Leu Asp Ser His Arg Gly Ala Val Phe
35 40 45
Ser Phe Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr Ala
50 55 60
Ile Val Asp Pro Pro Leu Gly Ile Ser Cys Phe Gly Arg Lys Met Trp
65 70 75 80
Ile Glu Ala Tyr Asn Gly Arg Gly Glu Val Leu Leu Asp Phe Ile Thr
85 90 95
Glu Lys Leu Lys Ala Thr Pro Asp Leu Thr Leu Gly Ala Ser Ser Thr
100 105 110
Arg Arg Leu Asp Leu Thr Val Asn Glu Pro Asp Arg Val Phe Thr Glu
115 120 125
Glu Glu Arg Ser Lys Ile Pro Thr Val Phe Thr Ala Leu Arg Ala Ile
130 135 140
Val Asp Leu Phe Tyr Ser Ser Ala Asp Ser Ala Ile Gly Leu Phe Gly
145 150 155 160
Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Lys Leu Ser
165 170 175
Leu Ala Arg Pro Glu Asp Gln Arg Asp Met Val Leu Phe Leu Pro Asp
180 185 190
Glu Ile Leu Val Val Asp His Tyr Ser Ala Lys Ala Trp Ile Asp Arg
195 200 205
Tyr Asp Phe Glu Lys Asp Gly Met Thr Thr Asp Gly Lys Ser Ser Asp
210 215 220
Ile Thr Pro Asp Pro Phe Lys Thr Thr Asp Thr Ile Pro Pro Lys Gly
225 230 235 240
Asp His Arg Pro Gly Glu Tyr Ser Glu Leu Val Val Lys Ala Lys Glu
245 250 255
Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys Phe
260 265 270
Met Glu Arg Cys Glu Ser Asn Pro Ser Ala Ile Ser Arg Arg Leu Lys
275 280 285
Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly Asp Gln
290 295 300
Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser Gly
305 310 315 320
Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly ThrIle Lys Arg Gly Asp
325 330 335
Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser Lys
340 345 350
Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn Asp
355 360 365
Lys Ser Arg Val Cys Glu Pro Gly Ser Val Lys Val Ile Gly Arg Arg
370 375 380
Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile Glu
385 390 395 400
Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser His
405 410 415
Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met Arg
420 425 430
Phe Ile Glu Gly His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly Ala
435 440 445
Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr Leu
450 455 460
Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly Ala
465 470 475 480
Thr Leu Leu Asn Asp Ser Asn Pro Gln Glu Glu Glu Ala Glu Thr Glu
485 490 495
Leu Lys Ala Ser Ala Met Ile Ser Ala Ile Arg Asp Ala Lys Gly Thr
500 505 510
Asn Ser Ala Ala Thr Lys Arg Asp Ala Ala Lys Val Gly Thr Gly Val
515 520 525
Lys Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu Ala
530 535 540
Asn Tyr Phe Arg Gln Thr Gly Ala Thr Val Ser Thr Val Arg Ser Pro
545 550 555 560
Val Ala Ala Asp Val Phe Asp Arg Phe Gln Pro Asp Leu Val Val Leu
565 570 575
Set Pro Gly Pro Gly Ser Pro Thr Asp Phe Asp Cys Lys Ala Thr Ile
580 585 590
Lys Ala Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu Gly
595 600 605
Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Glu Leu Arg Gln Leu Ala
610 615 620
Val Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro Gly
625 630 635 640
Leu Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr His
645 650 655
Ser Ile Phe Ala Asp Pro Ala Thr Leu Pro Arg Asp Phe Ile Ile Thr
660 665 670
Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ala Lys Glu
675 680 685
Pro Val Ala Ala Val Gln Phe His Pro Glu Set Ile Met Thr Leu Gly
690 695 700
Gln Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Val His Leu Thr
705 710 715 720
Arg Lys Ala Lys Thr Lys Ala Ala
725
<210>205
<211>2190
<212>DNA
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>205
atggtaacga tcattcagga tgacggagcg gagacctacg agacgaaagg cggcatccag 60
gtcagccgaa agcgccggcc caccgattat gccaacgcca tcgataatta catcgaaaag 120
cttgattccc atcgcggcgc ggttttttcg tgcaactatg aatatccggg ccgttacacc 180
cgctgggata cggccatcgt cgatccgccg ctcggcattt cctgttttgg ccgcaagatg 240
tggatcgaag cctataatgg ccgcggcgaa gtgctgctcg atttcattac ggaaaagctg 300
aaggcgacac ccgatctcac cctcggcgct tcctcgaccc gccggctcga tcttaccgtc 360
aacgaaccgg accgtgtctt caccgaagaa gaacgctcga aaatcccgac ggtcttcacc 420
gctctcagag ccatcgtcga cctcttctat tcgagcgcgg attcggccat cggcctgttc 480
ggtgccttcg gttacgatct cgccttccag ttcgacgcga tcaagctttc gctggcgcgt 540
ccggaagacc agcgtgacat ggtgctgttt ctgcccgatg aaatcctcgt cgttgatcac 600
tattccgcca aggcctggat cgaccgttac gatttcgaga aggacggcat gacgacggac 660
ggcaaatcct ccgacattac ccccgatccc ttcaagacca ccgataccat cccgcccaag 720
ggcgatcacc gtcccggcga atattccgag cttgtggtga aggccaagga aagcttccgc 780
cgcggcgacc tgttcgaggt cgttcccggc cagaaattca tggagcgttg cgaaagcaat 840
ccgtcggcga tttcccgccg cctgaaggcg atcaacccgt cgccctattc cttcttcatc 900
aatctcggcg atcaggaata tctggtcggc gcctcgccgg aaatgttcgt gcgcgtctcc 960
ggccgtcgca tcgagacctg cccgatatca ggcaccatca agcgcggcga cgatccgatt 1020
gccgacagcg agcagatttt gaaactgctc aactcgaaaa aggacgaatc cgaactgacc 1080
atgtgctcgg acgtggaccg caacgacaag agccgcgtct gcgagccggg ttcggtgaag 1140
gtcattggcc gccgccagat cgagatgtat tcacgcctca tccacaccgt cgatcacatc 1200
gaaggccgcc tgcgcgacga tatggacgcc tttgacggtt tcctcagcca cgcctgggcc 1260
gtcaccgtca ccggtgcacc aaagctgtgg gccatgcgct tcatcgaagg tcatgaaaag 1320
agcccgcgcg cctggtatgg cggtgcgatc ggcatggtcg gcttcaacgg cgacatgaat 1380
accggcctga cgctgcgcac catccggatc aaggacggta ttgccgaagt gcgcgccggc 1440
gcgaccctgc tcaatgattc caacccgcag gaagaagaag ccgaaaccga actgaaggcc 1500
tccgccatga tatcagccat tcgtgacgca aaaggcacca actctgccgc caccaagcgt 1560
gatgccgcca aagtcggcac cggcgtcaag atcctgctcg tcgaccacga agacagcttc 1620
gtgcacacgc tggcgaatta tttccgccag acgggcgcga cggtctcgac cgtcagatca 1680
ccggtcgcag ccgacgtgtt cgatcgcttc cagccggacc tcgttgtcct gtcgcccgga 1740
cccggcagcc cgacggattt cgactgcaag gcaacgatca aggccgcccg cgcccgcgat 1800
ctgccgatct tcggcgtttg cctcggtctg caggcattgg cagaagccta tggcggcgag 1860
ctgcgccagc ttgctgtgcc catgcacggc aagccttcgc gcatccgcgt gctggaaccc 1920
ggcctcgtct tctccggtct cggcaaggaa gtcacggtcg gtcgttacca ttcgatcttc 1980
gccgatcccg ccaccctgcc gcgtgatttc atcatcaccg cagaaagcga ggacggcacg 2040
atcatgggca tcgaacacgc caaggaaccg gtggccgccg ttcagttcca cccggaatcg 2100
atcatgacgc tcggacagga cgcgggcatg cggatgatcg agaatgtcgt ggtgcatctg 2160
acccgcaagg cgaagaccaa ggccgcgtga 2190
<210>206
<211>729
<212>PRT
<213>根癌土壤杆菌(Agrobacterium tumefaciens)
<400>206
Met Val Thr Ile Ile Gln Asp Asp Gly Ala Glu Thr Tyr Glu Thr Lys
1 5 10 15
Gly Gly Ile Gln Val Ser Arg Lys Arg Arg Pro Thr Asp Tyr Ala Asn
20 25 30
Ala Ile Asp Asn Tyr Ile Glu Lys Leu Asp Ser His Arg Gly Ala Val
35 40 45
Phe Ser Cys Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
50 55 60
Ala Ile Val Asp Pro Pro Leu Gly Ile Ser Cys Phe Gly Arg Lys Met
65 70 75 80
Trp Ile Glu Ala Tyr Asn Gly Arg Gly Glu Val Leu Leu Asp Phe Ile
85 90 95
Thr Glu Lys Leu Lys Ala Thr Pro Asp Leu Thr Leu Gly Ala Ser Ser
100 105 110
Thr Arg Arg Leu Asp Leu Thr Val Asn Glu Pro Asp Arg Val Phe Thr
115 120 125
Glu Glu Glu Arg Ser Lys Ile Pro Thr Val Phe Thr Ala Leu Arg Ala
130 135 140
Ile Val Asp Leu Phe Tyr Ser Ser Ala Asp Ser Ala Ile Gly Leu Phe
145 150 155 160
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Lys Leu
165 170 175
Ser Leu Ala Arg Pro Glu Asp Gln Arg Asp Met Val Leu Phe Leu Pro
180 185 190
Asp Glu Ile Leu Val Val Asp His Tyr Ser Ala Lys Ala Trp Ile Asp
195 200 205
Arg Tyr Asp Phe Glu Lys Asp Gly Met Thr Thr Asp Gly Lys Ser Ser
210 215 220
Asp Ile Thr Pro Asp Pro Phe Lys Thr Thr Asp Thr Ile Pro Pro Lys
225 230 235 240
Gly Asp His Arg Pro Gly Glu Tyr Ser Glu Leu Val Val Lys Ala Lys
245 250 255
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
260 265 270
Phe Met Glu Arg Cys Glu Ser Asn Pro Ser Ala Ile Ser Arg Arg Leu
275 280 285
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe PheIle Asn Leu Gly Asp
290 295 300
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
305 310 315 320
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
325 330 335
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
340 345 350
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
355 360 365
Asp Lys Ser Arg Val Cys Glu Pro Gly Ser Val Lys Val Ile Gly Arg
370 375 380
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
385 390 395 400
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
405 410 415
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
420 425 430
Arg Phe Ile Glu Gly His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
435 440 445
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
450 455 460
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
465 470 475 480
Ala Thr Leu Leu Asn Asp Ser Asn Pro Gln Glu Glu Glu Ala Glu Thr
485 490 495
Glu Leu Lys Ala Ser Ala Met Ile Ser Ala Ile Arg Asp Ala Lys Gly
500 505 510
Thr Asn Ser Ala Ala Thr Lys Arg Asp Ala Ala Lys Val Gly Thr Gly
515 520 525
Val Lys Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
530 535 540
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Thr Val Ser Thr Val Arg Ser
545 550 555 560
Pro Val Ala Ala Asp Val Phe Asp Arg Phe Gln Pro Asp Leu Val Val
565 570 575
Leu Ser Pro Gly Pro Gly Ser Pro Thr Asp Phe Asp Cys Lys Ala Thr
580 585 590
Ile Lys Ala Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
595 600 605
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Glu Leu Arg Gln Leu
610 615 620
Ala Val Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
625 630 635 640
Gly Leu Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
645 650 655
His Ser Ile Phe Ala Asp Pro Ala Thr Leu Pro Arg Asp Phe Ile Ile
660 665 670
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met GlyIle Glu His Ala Lys
675 680 685
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
690 695 700
Gly Gln Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Val His Leu
705 710 715 720
Thr Arg Lys Ala Lys Thr Lys Ala Ala
725
<210>207
<211>2190
<212>DNA
<213>人工序列
<220>
<223>来自根癌土壤杆菌(Agrobacterium tumefaciens)邻氨基苯甲酸合酶的密码子
最优化的S51C等位基因
<400>207
atggtgacca tcattcagga cgacggcgct gagacctacg agactaaggg cggtatccaa 60
gtgagccgta agcgtaggcc cactgactac gctaacgcca tcgacaacta catcgagaag 120
ctagactccc atcgcggcgc tgtgttctcc tgcaactacg aataccctgg gcgctacacg 180
aggtgggata ccgccatcgt cgatcctcca ttgggcatct cctgttttgg gcgtaagatg 240
tggatcgagg cgtacaacgg ccgtggcgaa gtcttgctgg acttcatcac ggagaagctc 300
aaggccacac cggacctcac cctcggcgct tcctcgaccc gccgcctcga ccttacggtc 360
aacgagccgg accgcgtgtt caccgaggaa gagcgtagca agatcccgac tgtcttcacc 420
gcgctcagag ccatcgtgga cctattctac tcttctgcgg acagcgccat cgggttgttc 480
ggtgccttcg gttacgacct cgcgttccag ttcgacgcca tcaagctctc gctcgcgcgg 540
ccggaggacc agcgagacat ggtgctcttc ctccctgacg agatcctggt cgtcgatcac 600
tattccgcga aggcgtggat cgaccggtac gacttcgaga aggatggcat gaccacggat 660
ggcaagagca gcgacatcac tcccgaccca ttcaagacca ccgacaccat ccctccaaag 720
ggcgatcacc gccctggcga gtattccgaa ctcgtggtga aggccaagga atccttccgg 780
cgcggcgatc tgtttgaggt ggttcccggc caaaagttca tggagaggtg cgagtcgaat 840
ccgtctgcga tcagtcgccg actgaaagcg atcaacccga gcccgtattc cttcttcatc 900
aacctcggcg atcaggaata tctggtcgga gcctcacccg agatgttcgt cagggtctcc 960
ggccgccgga tcgagacgtg cccaatttcc ggaaccatca agcgcggaga tgacccgata 1020
gccgactctg agcagatcct gaaactcttg aacagcaaga aggacgagtc cgagctgact 1080
atgtgctcag atgtggaccg aaacgacaag tcacgtgtct gcgagcccgg tagcgtcaag 1140
gtcattggcc gccgtcagat cgagatgtac tccaggctga ttcacacggt cgatcatatc 1200
gaagggcggc tgcgcgacga tatggacgca ttcgacgggt tcctcagtca cgcctgggcc 1260
gttactgtca ccggagcgcc taagctctgg gctatgaggt tcatcgaggg ccacgagaag 1320
agccctaggg cttggtatgg tggtgccatc ggcatggttg ggttcaacgg cgacatgaac 1380
accgggctga cgctccggac catcaggatc aaagacggca ttgccgaggt gagggccggt 1440
gccacgcttc tcaacgatag caaccctcag gaggaagagg cggagaccga gctgaaagcc 1500
tctgcgatga tctccgcgat tagagatgca aagggtacga acagtgctgc caccaagcgg 1560
gacgcagcca aggtgggcac cggcgtcaag attttacttg tcgatcacga ggactccttc 1620
gtgcacactc tggcgaacta cttccgccag acaggcgcga cggtctccac cgttaggtca 1680
ccggtggccg ctgacgtgtt cgataggttc cagcccgacc ttgtggtgct ctctcccggt 1740
cccggctcgc ccacggactt cgactgcaag gccaccatta aggccgccag ggccagggat 1800
ctgccaatct tcggcgtttg cctcgggctt caggcattgg ccgaggcata cggtggagag 1860
ctgaggcagc tcgccgtccc gatgcacggg aagccatccc gcatcagagt cctggagccc 1920
ggcctcgtct tctccggtct cgggaaggag gtcacggtcg gtcggtatca ttcgatcttc 1980
gccgatccgg caaccctccc gcgcgacttc atcataaccg ccgagtcgga ggacggaacg 2040
atcatgggaa tcgaacacgc caaggagccc gtagctgcgg ttcagttcca ccctgagtcc 2100
atcatgaccc tcggtcaaga tgcgggtatg cggatgatcg agaatgtggt ggttcacctc 2160
acccgcaagg ccaagaccaa ggcagcatga 2190
<210>208
<211>729
<212>PRT
<213>人工序列
<220>
<223>来自根癌土壤杆菌(Agrobacterium tumefaciens)邻氨基苯甲酸合酶的密码子
最优化的S51C等位基因
<400>208
Met Val Thr Ile Ile Gln Asp Asp Gly Ala Glu Thr Tyr Glu Thr Lys
1 5 10 15
Gly Gly Ile Gln Val Ser Arg Lys Arg Arg Pro Thr Asp Tyr Ala Asn
20 25 30
Ala Ile Asp Asn Tyr Ile Glu Lys Leu Asp Ser His Arg Gly Ala Val
35 40 45
Phe Ser Cys Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
50 55 60
Ala Ile Val Asp Pro Pro Leu Gly Ile Ser Cys Phe Gly Arg Lys Met
65 70 75 80
Trp Ile Glu Ala Tyr Asn Gly Arg Gly Glu Val Leu Leu Asp Phe Ile
85 90 95
Thr Glu Lys Leu Lys Ala Thr Pro Asp Leu Thr Leu Gly Ala Ser Ser
100 105 110
Thr Arg Arg Leu Asp Leu Thr Val Asn Glu Pro Asp Arg Val Phe Thr
115 120 125
Glu Glu Glu Arg Ser Lys Ile Pro Thr Val Phe Thr Ala Leu Arg Ala
130 135 140
Ile Val Asp Leu Phe Tyr Ser Ser Ala Asp Ser Ala Ile Gly Leu Phe
145 150 155 160
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Lys Leu
165 170 175
Ser Leu Ala Arg Pro Glu Asp Gln Arg Asp Met Val Leu Phe Leu Pro
180 185 190
Asp Glu Ile Leu Val Val Asp His Tyr Ser Ala Lys Ala Trp Ile Asp
195 200 205
Arg Tyr Asp Phe Glu Lys Asp Gly Met Thr Thr Asp Gly Lys Ser Ser
210 215 220
Asp Ile Thr Pro Asp Pro Phe Lys Thr Thr Asp Thr Ile Pro Pro Lys
225 230 235 240
Gly Asp His Arg Pro Gly Glu Tyr Ser Glu Leu Val Val Lys Ala Lys
245 250 255
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
260 265 270
Phe Met Glu Arg Cys Glu Ser Asn Pro Ser Ala Ile Ser Arg Arg Leu
275 280 285
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly Asp
290 295 300
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
305 310 315 320
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
325 330 335
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
340 345 350
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
355 360 365
Asp Lys Ser Arg Val Cys Glu Pro Gly Ser Val Lys Val Ile Gly Arg
370 375 380
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
385 390 395 400
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
405 410 415
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
420 425 430
Arg Phe Ile Glu Gly His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
435 440 445
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
450 455 460
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
465 470 475 480
Ala Thr Leu Leu Asn Asp Ser Asn Pro Gln Glu Glu Glu Ala Glu Thr
485 490 495
Glu Leu Lys Ala Ser Ala Met Ile Ser Ala Ile Arg Asp Ala Lys Gly
500 505 510
Thr Asn Ser Ala Ala Thr Lys Arg Asp Ala Ala Lys Val Gly Thr Gly
515 520 525
Val Lys Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
530 535 540
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Thr Val Ser Thr Val Arg Ser
545 550 555 560
Pro Val Ala Ala Asp Val Phe Asp Arg Phe Gln Pro Asp Leu Val Val
565 570 575
Leu Ser Pro Gly Pro Gly Ser Pro Thr Asp Phe Asp Cys Lys Ala Thr
580 585 590
Ile Lys Ala Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
595 600 605
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Glu Leu Arg Gln Leu
610 615 620
Ala Val Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
625 630 635 640
Gly Leu Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
645 650 655
His Ser Ile Phe Ala Asp Pro Ala Thr Leu Pro Arg Asp Phe Ile Ile
660 665 670
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ala Lys
675 680 685
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
690 695 700
Gly Gln Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Val His Leu
705 710 715 720
Thr Arg Lys Ala Lys Thr Lys Ala Ala
725
<210>209
<211>2191
<212>DNA
<213>苜蓿中华根瘤菌(Sinorhizobium meliloti)
<400>209
atggcagcgg taattctgga agacggcgcg gagagttata ccacgaaggg tggcatcgtc 60
gtcacccgca ggcggcgtga ggcatcctac agcgacgcga tcgccggtta tgtcgaccgg 120
ctggacgaac gccgcggcgc ggtcttttcc tcgaactacg aatatcccgg ccgctatacc 180
cgctgggaca ctgcggtggt cgacccgccg cttgccatct cctccttcgg tcgctcgctc 240
tggatcgaag cctataacga acgcggcgaa gtgctgctgg cgctgatcgc cgaggatctg 300
aagtccgttg ccgacatcac gctcggctca cttgccgccc gccgcctcga cctcaccatc 360
aacgagcccg atcgtgtctt caccgaggaa gagcggtcga agatgccgac ggtctttacg 420
gttcttcgcg cggtgacgaa cctcttccac tcggaggagg actcgaacct cggcctctat 480
ggcgccttcg gctacgacct cgccttccag ttcgatgcga tcgaactgaa gctttcgcgt 540
ccggacgacc agcgcgacat ggttctcttt ctgccggacg agatccttgt ggtcgatcac 600
tatgcggcca aggcctggat cgaccgctac gatttcgcca gggagaacct ttcgaccgag 660
ggcaaggcag cggacattgc tcccgagccg ttccgcagcg tcgacagcat cccgccgcac 720
ggggatcacc gcccgggcga atatgccgag ctcgtcgtca aggcgaagga aagcttccgt 780
cgcggcgatc ttttcgaagt ggtgccgggg cagaaattct acgagcgctg cgaaagccgc 840
ccgtccgaga tttccaaccg gctgaaggcg atcaatccgt cgccctattc cttcttcatc 900
aatctcggca accaggaata tctcgtcggt gcttcgccgg agatgttcgt gcgcgtttcc 960
ggccggcgca tcgagacctg cccgatctcc ggtacgatca agcgcggcga cgatccgatc 1020
gccgacagcg agcagatcct gaagctcttg aactcgaaga aggacgagtc cgagctcacc 1080
atgtgctcgg acgtcgaccg caacgacaag agccgggtct gcgtgccggg ctcggtcaag 1140
gtgatcggcc ggcgtcagat cgagatgtat tcgcggctga tccacacggt cgatcacatc 1200
gaggggcgcc tgcgcgacga tatggacgcc ttcgacgggt tcctcagcca cgcctgggcg 1260
gtgaccgtta ccggcgcgcc aaagctctgg gccatgcgct tcatcgagag ccacgagaag 1320
agcccgcgtg cctggtatgg cggcgcgatc ggcatggtcg gcttcaacgg cgacatgaat 1380
accgggctga ccttgcgtac catccgcatc aaggacggga tcgccgaggt gagggcgggt 1440
gcgacgctcc tctatgattc caatccggaa gaagaagaag ccgaaaccga actgaaggcc 1500
tctgccatga ttgcagccat ccgcgacgcg aaatccgcaa acagcgccaa atccgcgcgc 1560
gatgtcgccg ccgtcggcgc cggagtcagc atcctgctcg tcgatcacga ggacagcttc 1620
gtccataccc tcgcgaacta cttccgccag accggcgcgt ccgtcaccac cgtgcgcacg 1680
ccggtggccg aggaaatctt cgaccgggtc aagccggacc tcgtcgtgct ttcgcccggt 1740
cccggcaccc cgaaggactt cgactgcaag gcgacgatca agaaggcgcg ggcgcgggac 1800
ctgccgatct tcggcgtctg cctggggctg caggcgctcg cggaggccta tggcggcgac 1860
cttcgtcaac tggcgatccc gatgcatggg aagccctcgc gcatccgcgt gctcgaaccc 1920
ggcatcgtct tctccggcct cggcaaggag gtgacggtcg ggcgctatca ttcgattttc 1980
gccgatccgt ccaacctgcc gcgcgaattc gtgatcacgg ccgaaagcga agatggtacg 2040
atcatgggca tcgaacacag caaggagccg gtggcggccg tgcagttcca tccggaatcg 2100
atcatgacgc tgggcggcga cgccggcatg cggatgatcg agaacgtggt tgcccatctc 2160
gccaagcggg cgaagaccaa ggcagcctga a 2191
<210>210
<211>729
<212>PRT
<213>苜蓿中华根瘤菌(Sinorhizobium meliloti)
<400>210
Met Ala Ala Val Ile Leu Glu Asp Gly Ala Glu Ser Tyr Thr Thr Lys
1 5 10 15
Gly Gly Ile Val Val Thr Arg Arg Arg Arg Glu Ala Ser Tyr Ser Asp
20 25 30
Ala Ile Ala Gly Tyr Val Asp Arg Leu Asp Glu Arg Arg Gly Ala Val
35 40 45
Phe Ser Ser Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
50 55 60
Ala Val Val Asp Pro Pro Leu Ala Ile Ser Ser Phe Gly Arg Ser Leu
65 70 75 80
Trp Ile Glu Ala Tyr Asn Glu Arg Gly Glu Val Leu Leu Ala Leu Ile
85 90 95
Ala Glu Asp Leu Lys Ser Val Ala Asp Ile Thr Leu Gly Ser Leu Ala
100 105 110
Ala Arg Arg Leu Asp Leu Thr Ile Asn Glu Pro Asp Arg Val Phe Thr
115 120 125
Glu Glu Glu Arg Ser Lys Met Pro Thr Val Phe Thr Val Leu Arg Ala
130 135 140
Val Thr Asn Leu Phe His Ser Glu Glu Asp Ser Asn Leu Gly Leu Tyr
145 150 155 160
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Glu Leu
165 170 175
Lys Leu Ser Arg Pro Asp Asp Gln Arg Asp Met Val Leu Phe Leu Pro
180 185 190
Asp Glu Ile Leu Val Val Asp His Tyr Ala Ala Lys Ala Trp Ile Asp
195 200 205
Arg Tyr Asp Phe Ala Arg Glu Asn Leu Ser Thr Glu Gly Lys Ala Ala
210 215 220
Asp Ile Ala Pro Glu Pro Phe Arg Ser Val Asp Ser Ile Pro Pro His
225 230 235 240
Gly Asp His Arg Pro Gly Glu Tyr Ala Glu Leu Val Val Lys Ala Lys
245 250 255
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
260 265 270
Phe Tyr Glu Arg Cys Glu Ser Arg Pro Ser Glu Ile Ser Asn Arg Leu
275 280 285
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly Asn
290 295 300
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
305 310 315 320
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
325 330 335
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
340 345 350
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
355 360 365
Asp Lys Ser Arg Val Cys Val Pro Gly Ser Val Lys Val Ile Gly Arg
370 375 380
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
385 390 395 400
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
405 410 415
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
420 425 430
Arg Phe Ile Glu Ser His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
435 440 445
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
450 455 460
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
465 470 475 480
Ala Thr Leu Leu Tyr Asp Ser Asn Pro Glu Glu Glu Glu Ala Glu Thr
485 490 495
Glu Leu Lys Ala Ser Ala Met Ile Ala Ala Ile Arg Asp Ala Lys Ser
500 505 510
Ala Asn Ser Ala Lys Ser Ala Arg Asp Val Ala Ala Val Gly Ala Gly
515 520 525
Val Ser Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
530 535 540
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Ser Val Thr Thr Val Arg Thr
545 550 555 560
Pro Val Ala Glu Glu Ile Phe Asp Arg Val Lys Pro Asp Leu Val Val
565 570 575
Leu Ser Pro Gly Pro Gly Thr Pro Lys Asp Phe Asp Cys Lys Ala Thr
580 585 590
Ile Lys Lys Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
595 600 605
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Asp Leu Arg Gln Leu
610 615 620
Ala Ile Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
625 630 635 640
Gly Ile Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
645 650 655
His Ser Ile Phe Ala Asp Pro Ser Asn Leu Pro Arg Glu Phe Val Ile
660 665 670
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ser Lys
675 680 685
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
690 695 700
Gly Gly Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Ala His Leu
705 710 715 720
Ala Lys Arg Ala Lys Thr Lys Ala Ala
725
<210>211
<211>2190
<212>DNA
<213>苜蓿中华根瘤菌(Sinorhizobium meliloti)
<400>211
atggcagcgg taattctgga agacggcgcg gagagttata ccacgaaggg tggcatcgtc 60
gtcacccgca ggcggcgtga ggcatcctac agcgacgcga tcgccggtta tgtcgaccgg 120
ctggacgaac gccgcggcgc ggtcttttcc tgcaactacg aatatcccgg ccgctatacc 180
cgctgggaca ctgcggtggt cgacccgccg cttgccatct cctccttcgg tcgctcgctc 240
tggatcgaag cctataacga acgcggcgaa gtgctgctgg cgctgatcgc cgaggatctg 300
aagtccgttg ccgacatcac gctcggctca cttgccgccc gccgcctcga cctcaccatc 360
aacgagcccg atcgtgtctt caccgaggaa gagcggtcga agatgccgac ggtctttacg 420
gttcttcgcg cggtgacgaa cctcttccac tcggaggagg actcgaacct cggcctctat 480
ggcgccttcg gctacgacct cgccttccag ttcgatgcga tcgaactgaa gctttcgcgt 540
ccggacgacc agcgcgacat ggttctcttt ctgccggacg agatccttgt ggtcgatcac 600
tatgcggcca aggcctggat cgaccgctac gatttcgcca gggagaacct ttcgaccgag 660
ggcaaggcag cggacattgc tcccgagccg ttccgcagcg tcgacagcat cccgccgcac 720
ggggatcacc gcccgggcga atatgccgag ctcgtcgtca aggcgaagga aagcttccgt 780
cgcggcgatc ttttcgaagt ggtgccgggg cagaaattct acgagcgctg cgaaagccgc 840
ccgtccgaga tttccaaccg gctgaaggcg atcaatccgt cgccctattc cttcttcatc 900
aatctcggca accaggaata tctcgtcggt gcttcgccgg agatgttcgt gcgcgtttcc 960
ggccggcgca tcgagacctg cccgatctcc ggtacgatca agcgcggcga cgatccgatc 1020
gccgacagcg agcagatcct gaagctcttg aactcgaaga aggacgagtc cgagctcacc 1080
atgtgctcgg acgtcgaccg caacgacaag agccgggtct gcgtgccggg ctcggtcaag 1140
gtgatcggcc ggcgtcagat cgagatgtat tcgcggctga tccacacggt cgatcacatc 1200
gaggggcgcc tgcgcgacga tatggacgcc ttcgacgggt tcctcagcca cgcctgggcg 1260
gtgaccgtta ccggcgcgcc aaagctctgg gccatgcgct tcatcgagag ccacgagaag 1320
agcccgcgtg cctggtatgg cggcgcgatc ggcatggtcg gcttcaacgg cgacatgaat 1380
accgggctga ccttgcgtac catccgcatc aaggacggga tcgccgaggt gagggcgggt 1440
gcgacgctcc tctatgattc caatccggaa gaagaagaag ccgaaaccga actgaaggcc 1500
tctgccatga ttgcagccat ccgcgacgcg aaatccgcaa acagcgccaa atccgcgcgc 1560
gatgtcgccg ccgtcggcgc cggagtcagc atcctgctcg tcgatcacga ggacagcttc 1620
gtccataccc tcgcgaacta cttccgccag accggcgcgt ccgtcaccac cgtgcgcacg 1680
ccggtggccg aggaaatctt cgaccgggtc aagccggacc tcgtcgtgct ttcgcccggt 1740
cccggcaccc cgaaggactt cgactgcaag gcgacgatca agaaggcgcg ggcgcgggac 1800
ctgccgatct tcggcgtctg cctggggctg caggcgctcg cggaggccta tggcggcgac 1860
cttcgtcaac tggcgatccc gatgcatggg aagccctcgc gcatccgcgt gctcgaaccc 1920
ggcatcgtct tctccggcct cggcaaggag gtgacggtcg ggcgctatca ttcgattttc 1980
gccgatccgt ccaacctgcc gcgcgaattc gtgatcacgg ccgaaagcga agatggtacg 2040
atcatgggca tcgaacacag caaggagccg gtggcggccg tgcagttcca tccggaatcg 2100
atcatgacgc tgggcggcga cgccggcatg cggatgatcg agaacgtggt tgcccatctc 2160
gccaagcggg cgaagaccaa ggcagcctga 2190
<210>212
<211>729
<212>PRT
<213>苜蓿中华根瘤菌(Sinorhizobium meliloti)
<400>212
Met Ala Ala Val Ile Leu Glu Asp Gly Ala Glu Ser Tyr Thr Thr Lys
1 5 10 15
Gly Gly Ile Val Val Thr Arg Arg Arg Arg Glu Ala Ser Tyr Ser Asp
20 25 30
Ala Ile Ala Gly Tyr Val Asp Arg Leu Asp Glu Arg Arg Gly Ala Val
35 40 45
Phe Ser Cys Asn Tyr Glu Tyr Pro Gly Arg Tyr Thr Arg Trp Asp Thr
50 55 60
Ala Val Val Asp Pro Pro Leu Ala Ile Ser Ser Phe Gly Arg Ser Leu
65 70 75 80
Trp Ile Glu Ala Tyr Asn Glu Arg Gly Glu Val Leu Leu Ala Leu Ile
85 90 95
Ala Glu Asp Leu Lys Ser Val Ala Asp Ile Thr Leu Gly Ser Leu Ala
100 105 110
Ala Arg Arg Leu Asp Leu Thr Ile Asn Glu Pro Asp Arg Val Phe Thr
115 120 125
Glu Glu Glu Arg Ser Lys Met Pro Thr Val Phe Thr Val Leu Arg Ala
130 135 140
Val Thr Asn Leu Phe His Ser Glu Glu Asp Ser Asn Leu Gly Leu Tyr
145 150 155 160
Gly Ala Phe Gly Tyr Asp Leu Ala Phe Gln Phe Asp Ala Ile Glu Leu
165 170 175
Lys Leu Ser Arg Pro Asp Asp Gln Arg Asp Met Val Leu Phe Leu Pro
180 185 190
Asp Glu Ile Leu Val Val Asp His Tyr Ala Ala Lys Ala Trp Ile Asp
195 200 205
Arg Tyr Asp Phe Ala Arg Glu Asn Leu Ser Thr Glu Gly Lys Ala Ala
210 215 220
Asp Ile Ala Pro Glu Pro Phe Arg Ser Val Asp Ser Ile Pro Pro His
225 230 235 240
Gly Asp His Arg Pro Gly Glu Tyr Ala Glu Leu Val Val Lys Ala Lys
245 250 255
Glu Ser Phe Arg Arg Gly Asp Leu Phe Glu Val Val Pro Gly Gln Lys
260 265 270
Phe Tyr Glu Arg Cys Glu Ser Arg Pro Ser Glu Ile Ser Asn Arg Leu
275 280 285
Lys Ala Ile Asn Pro Ser Pro Tyr Ser Phe Phe Ile Asn Leu Gly Asn
290 295 300
Gln Glu Tyr Leu Val Gly Ala Ser Pro Glu Met Phe Val Arg Val Ser
305 310 315 320
Gly Arg Arg Ile Glu Thr Cys Pro Ile Ser Gly Thr Ile Lys Arg Gly
325 330 335
Asp Asp Pro Ile Ala Asp Ser Glu Gln Ile Leu Lys Leu Leu Asn Ser
340 345 350
Lys Lys Asp Glu Ser Glu Leu Thr Met Cys Ser Asp Val Asp Arg Asn
355 360 365
Asp Lys Ser Arg Val Cys Val Pro Gly Ser Val Lys Val Ile Gly Arg
370 375 380
Arg Gln Ile Glu Met Tyr Ser Arg Leu Ile His Thr Val Asp His Ile
385 390 395 400
Glu Gly Arg Leu Arg Asp Asp Met Asp Ala Phe Asp Gly Phe Leu Ser
405 410 415
His Ala Trp Ala Val Thr Val Thr Gly Ala Pro Lys Leu Trp Ala Met
420 425 430
Arg Phe Ile Glu Ser His Glu Lys Ser Pro Arg Ala Trp Tyr Gly Gly
435 440 445
Ala Ile Gly Met Val Gly Phe Asn Gly Asp Met Asn Thr Gly Leu Thr
450 455 460
Leu Arg Thr Ile Arg Ile Lys Asp Gly Ile Ala Glu Val Arg Ala Gly
465 470 475 480
Ala Thr Leu Leu Tyr Asp Ser Asn Pro Glu Glu Glu Glu Ala Glu Thr
485 490 495
Glu Leu Lys Ala Ser Ala Met Ile Ala Ala Ile Arg Asp Ala Lys Ser
500 505 510
Ala Asn Ser Ala Lys Ser Ala Arg Asp Val Ala Ala Val Gly Ala Gly
515 520 525
Val Ser Ile Leu Leu Val Asp His Glu Asp Ser Phe Val His Thr Leu
530 535 540
Ala Asn Tyr Phe Arg Gln Thr Gly Ala Ser Val Thr Thr Val Arg Thr
545 550 555 560
Pro Val Ala Glu Glu Ile Phe Asp Arg Val Lys Pro Asp Leu Val Val
565 570 575
Leu Ser Pro Gly Pro Gly Thr Pro Lys Asp Phe Asp Cys Lys Ala Thr
580 585 590
Ile Lys Lys Ala Arg Ala Arg Asp Leu Pro Ile Phe Gly Val Cys Leu
595 600 605
Gly Leu Gln Ala Leu Ala Glu Ala Tyr Gly Gly Asp Leu Arg Gln Leu
610 615 620
Ala Ile Pro Met His Gly Lys Pro Ser Arg Ile Arg Val Leu Glu Pro
625 630 635 640
Gly Ile Val Phe Ser Gly Leu Gly Lys Glu Val Thr Val Gly Arg Tyr
645 650 655
His Ser Ile Phe Ala Asp Pro Ser Asn Leu Pro Arg Glu Phe Val Ile
660 665 670
Thr Ala Glu Ser Glu Asp Gly Thr Ile Met Gly Ile Glu His Ser Lys
675 680 685
Glu Pro Val Ala Ala Val Gln Phe His Pro Glu Ser Ile Met Thr Leu
690 695 700
Gly Gly Asp Ala Gly Met Arg Met Ile Glu Asn Val Val Ala His Leu
705 710 715 720
Ala Lys Arg Ala Lys Thr Lys Ala Ala
725
<210>213
<211>13596
<212>DNA
<213>人工序列
<220>
<223>pMON68065;图48;Zm-ASA2-CTP+18::AgroAS(S51C)突变等位基因的表达载体
<400>213
gcgccaaatc gtgaagtttc tcatctaagc ccccatttgg acgtgaatgt agacacgtcg 60
aaataaagat ttccgaatta gaataatttg tttattgctt tcgcctataa atacgacgga 120
tcgtaatttg tcgttttatc aaaatgtact ttcattttat aataacgctg cggacatcta 180
catttttgaa ttgaaaaaaa attggtaatt actctttctt tttctccata ttgaccatca 240
tactcattgc tgatccatgt agatttcccg gacatgaagc catttacaat tgaatatatc 300
ctgccgccgc tgccgctttg cacccggtgg agcttgcatg ttggtttcta cgcagaactg 360
agccggttag gcagataatt tccattgaga actgagccat gtgcaccttc cccccaacac 420
ggtgagcgac ggggcaacgg agtgatccac atgggacttt tcctagcttg gctgccattt 480
ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt 540
tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg 600
cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa 660
aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg 720
cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc 780
tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac 840
cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag 900
gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc 960
ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct 1020
catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccggccgcg 1080
gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc 1140
cagcccagcg gcgagggcaa ccagcccggt gagcgtcgga aagggtcgat cgaccgatgc 1200
ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg 1260
ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct 1320
gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg 1380
cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac 1440
gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct 1500
tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg 1560
gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc 1620
agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc 1680
tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg 1740
taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg 1800
ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat 1860
tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat 1920
atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc 1980
ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg 2040
ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg 2100
ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta 2160
aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg 2220
atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc 2280
ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg 2340
ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt 2400
catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa 2460
acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct 2520
ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga 2580
ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct 2640
ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 2700
acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca 2760
gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta 2820
ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 2880
atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 2940
cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 3000
gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 3060
ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 3120
agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 3180
tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 3240
ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 3300
gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 3360
ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 3420
gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 3480
aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 3540
aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 3600
ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 3660
gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 3720
gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 3780
tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 3840
ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 3900
ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 3960
atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 4020
ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 4080
tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 4140
attgctgcag gtcgggagca caggatgacg cctaacaatt cattcaagcc gacaccgctt 4200
cgcggcgcgg cttaattcag gagttaaaca tcatgaggga agcggtgatc gccgaagtat 4260
cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg 4320
ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt 4380
tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc 4440
ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca 4500
ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg 4560
gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg 4620
atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg 4680
cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct 4740
taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt 4800
tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg 4860
actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctaggcagg 4920
cttatcttgg acaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgttc 4980
actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctaaca attcgttcaa 5040
gccgacgccg cttcgcggcg cggcttaact caagcgttag atgctgcagg catcgtggtg 5100
tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 5160
acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgaggat 5220
ttttcggcgc tgcgctacgt ccgcgaccgc gttgagggat caagccacag cagcccactc 5280
gaccttctag ccgacccaga cgagccaagg gatctttttg gaatgctgct ccgtcgtcag 5340
gctttccgac gtttgggtgg ttgaacagaa gtcattatcg cacggaatgc caagcactcc 5400
cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac 5460
gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa 5520
tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatcccca 5580
tcaagctagc ttctgcaggt cctgctcgag gtcactaagc aactaacttt gaggaatgag 5640
gtgatgatga attaactcac tccattccac aaaccaaaca aaaatttgag gagtgagaag 5700
atgattgact atctcattcc tcaaaccaaa cacctcaaat atatctgcta tcgggattgg 5760
cattcctgta tccctacgcc cgtgtacccc ctgtttagag aacctccaaa ggtataagat 5820
ggcgaagatt attgttgtct tgtctttcat catatatcga gtctttccct aggatattat 5880
tattggcaat gagcattaca cggttaatcg attgagagaa catgcatctc accttcagca 5940
aataattacg ataatccata ttttacgctt cgtaacttct catgagtttc gatatacaaa 6000
tttgttttct ggacacccta ccattcatcc tcttcggaga agagaggaag tgtcctcaat 6060
ttaaatatgt tgtcatgctg tagttcttca caaaatctca acaggtacca agcacattgt 6120
ttccacaaat tatattttag tcacaataaa tctatattat tattaatata ctaaaactat 6180
actgacgctc agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc 6240
agaaaatagt gagacacgga agacaaaaga agtaaaagag gcccggacta cggcccacat 6300
gagattcggc cccgccacct ccggcaacca gcggccgatc caacggcagt gcgcgcacac 6360
acacaacctc gtatatatcg ccgcgcggaa gcggcgcgac cgaggaagcc ttgtcctcga 6420
caccccctac acaggtgtcg cgctgccccc gacacgagtc ccgcatgcgt cccacgcggc 6480
cgcgccagat cccgcctccg cgcgttgcca cgccctctat aaacacccag ctctccctcg 6540
ccctcatcta cctcactcgt agtcgtagct caagcatcag cggcagcggc agcggcagga 6600
tctctgggca gcgtgcgcac gtggggtatc tagctcgctc tgctagccta ccaatcgaat 6660
tcctgcaggt cgactctaga ggatctaccg tcttcggtac gcgctcactc cgccctctgc 6720
ctttgttact gccacgtttc tctgaatgct ctcttgtgtg gtgattgctg agagtggttt 6780
agctggatct agaattacac tctgaaatcg tgttctgcct gtgctgatta cttgccgtcc 6840
tttgtagcag caaaatatag ggacatggta gtacgaaacg aagatagaac ctacacagca 6900
atacgagaaa tgtgtaattt ggtgcttagc ggtatttatt taagcacatg ttggtgttat 6960
agggcacttg gattcagaag tttgctgtta atttaggcac aggcttcata ctacatgggt 7020
caatagtata gggattcata ttataggcga tactataata atttgttcgt ctgcagagct 7080
tattatttgc caaaattaga tattcctatt ctgtttttgt ttgtgtgctg ttaaattgtt 7140
aacgcctgaa ggaataaata taaatgacga aattttgatg tttatctctg ctcctttatt 7200
gtgaccataa gtcaagatca gatgcacttg ttttaaatat tgttgtctga agaaataagt 7260
actgacagta ttttgatgca ttgatctgct tgtttgttgt aacaaaattt aaaaataaag 7320
agtttccttt ttgttgctct ccttacctcc tgatggtatc tagtatctac caactgacac 7380
tatattgctt ctctttacat acgtatcttg ctcgatgcct tctccctagt gttgaccagt 7440
gttactcaca tagtctttgc tcatttcatt gtaatgcaga taccaagcgg cctctagagg 7500
actccgatct atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt 7560
cccggcggcg cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag 7620
ccggagcgga accagcgggg tgaaatgctc tgctgccgtg acgccgcagg cgagcccagt 7680
gattagcagg agcgctgcgg cggccatggt aacgatcatt caggatgacg gagcggagac 7740
ctacgagacg aaaggcggca tccaggtcag ccgaaagcgc cggcccaccg attatgccaa 7800
cgccatcgat aattacatcg aaaagcttga ttcccatcgc ggcgcggttt tttcgtgcaa 7860
ctatgaatat ccgggccgtt acacccgctg ggatacggcc atcgtcgatc cgccgctcgg 7920
catttcctgt tttggccgca agatgtggat cgaagcctat aatggccgcg gcgaagtgct 7980
gctcgatttc attacggaaa agctgaaggc gacacccgat ctcaccctcg gcgcttcctc 8040
gacccgccgg ctcgatctta ccgtcaacga accggaccgt gtcttcaccg aagaagaacg 8100
ctcgaaaatc ccgacggtct tcaccgctct cagagccatc gtcgacctct tctattcgag 8160
cgcggattcg gccatcggcc tgttcggtgc cttcggttac gatctcgcct tccagttcga 8220
cgcgatcaag ctttcgctgg cgcgtccgga agaccagcgt gacatggtgc tgtttctgcc 8280
cgatgaaatc ctcgtcgttg atcactattc cgccaaggcc tggatcgacc gttacgattt 8340
cgagaaggac ggcatgacga cggacggcaa atcctccgac attacccccg atcccttcaa 8400
gaccaccgat accatcccgc ccaagggcga tcaccgtccc ggcgaatatt ccgagcttgt 8460
ggtgaaggcc aaggaaagct tccgccgcgg cgacctgttc gaggtcgttc ccggccagaa 8520
attcatggag cgttgcgaaa gcaatccgtc ggcgatttcc cgccgcctga aggcgatcaa 8580
cccgtcgccc tattccttct tcatcaatct cggcgatcag gaatatctgg tcggcgcctc 8640
gccggaaatg ttcgtgcgcg tctccggccg tcgcatcgag acctgcccga tatcaggcac 8700
catcaagcgc ggcgacgatc cgattgccga cagcgagcag attttgaaac tgctcaactc 8760
gaaaaaggac gaatccgaac tgaccatgtg ctcggacgtg gaccgcaacg acaagagccg 8820
cgtctgcgag ccgggttcgg tgaaggtcat tggccgccgc cagatcgaga tgtattcacg 8880
cctcatccac accgtcgatc acatcgaagg ccgcctgcgc gacgatatgg acgcctttga 8940
cggtttcctc agccacgcct gggccgtcac cgtcaccggt gcaccaaagc tgtgggccat 9000
gcgcttcatc gaaggtcatg aaaagagccc gcgcgcctgg tatggcggtg cgatcggcat 9060
ggtcggcttc aacggcgaca tgaataccgg cctgacgctg cgcaccatcc ggatcaagga 9120
cggtattgcc gaagtgcgcg ccggcgcgac cctgctcaat gattccaacc cgcaggaaga 9180
agaagccgaa accgaactga aggcctccgc catgatatca gccattcgtg acgcaaaagg 9240
caccaactct gccgccacca agcgtgatgc cgccaaagtc ggcaccggcg tcaagatcct 9300
gctcgtcgac cacgaagaca gcttcgtgca cacgctggcg aattatttcc gccagacggg 9360
cgcgacggtc tcgaccgtca gatcaccggt cgcagccgac gtgttcgatc gcttccagcc 9420
ggacctcgtt gtcctgtcgc ccggacccgg cagcccgacg gatttcgact gcaaggcaac 9480
gatcaaggcc gcccgcgccc gcgatctgcc gatcttcggc gtttgcctcg gtctgcaggc 9540
attggcagaa gcctatggcg gcgagctgcg ccagcttgct gtgcccatgc acggcaagcc 9600
ttcgcgcatc cgcgtgctgg aacccggcct cgtcttctcc ggtctcggca aggaagtcac 9660
ggtcggtcgt taccattcga tcttcgccga tcccgccacc ctgccgcgtg atttcatcat 9720
caccgcagaa agcgaggacg gcacgatcat gggcatcgaa cacgccaagg aaccggtggc 9780
cgccgttcag ttccacccgg aatcgatcat gacgctcgga caggacgcgg gcatgcggat 9840
gatcgagaat gtcgtggtgc atctgacccg caaggcgaag accaaggccg cgtgatggcg 9900
ctcgatgaca cggttatatc actagtgcgg ccatcggatc cgttggcaat gcggataaag 9960
aataactaaa taaataaata aataaattgc aagcaattgc gttgctgcta tgtactgtaa 10020
aagtttctta taatatcagt tctgaatgct aaggacatcc ctcaagatgg tctttctatt 10080
tttgtgttcc cgttccaatg tactgttggt atcctcttgg agattcatca atatgagaaa 10140
acagagaatg gacaaccctc ccttatctta tggctcgagc ggccgctcta gaactagtgg 10200
atccccccct taattaaggg ggctgcagga attcataact tcgtataatg tatgctatac 10260
gaagttatgt ttcgaggtca ttcatatgct tgagaagaga gtcgggatag tccaaaataa 10320
aacaaaggta agattacctg gtcaaaagtg aaaacatcag ttaaaaggtg gtataaagta 10380
aaatatcggt aataaaaggt ggcccaaagt gaaatttact cttttctact attataaaaa 10440
ttgaggatgt ttttgtcggt actttgatac gtcatttttg tatgaattgg tttttaagtt 10500
tattcgcttt tggaaatgca tatctgtatt tgagtcgggt tttaagttcg tttgcttttg 10560
taaatacaga gggatttgta taagaaatat ctttagaaaa acccatatgc taatttgaca 10620
taatttttga gaaaaatata tattcaggcg aattctcaca atgaacaata ataagattaa 10680
aatagctttc ccccgttgca gcgcatgggt attttttcta gtaaaaataa aagataaact 10740
tagactcaaa acatttacaa aaacaacccc taaagttcct aaagcccaaa gtgctatcca 10800
cgatccatag caagcccagc ccaacccaac ccaacccaac ccaccccagt ccagccaact 10860
ggacaatagt ctccacaccc ccccactatc accgtgagtt gtccgcacgc accgcacgtc 10920
tcgcagccaa aaaaaaaaag aaagaaaaaa aagaaaaaga aaaaacagca ggtgggtccg 10980
ggtcgtgggg gccggaaacg cgaggaggat cgcgagccag cgacgaggcc ggccctccct 11040
ccgcttccaa agaaacgccc cccatcgcca ctatatacat acccccccct ctcctcccat 11100
ccccccaacc ctaccaccac caccaccacc acctccacct cctcccccct cgctgccgga 11160
cgacgagctc ctcccccctc cccctccgcc gccgccgcgc cggtaaccac cccgcccctc 11220
tcctctttct ttctccgttt ttttttccgt ctcggtctcg atctttggcc ttggtagttt 11280
gggtgggcga gaggcggctt cgtgcgcgcc cagatcggtg cgcgggaggg gcgggatctc 11340
gcggctgggg ctctcgccgg cgtggatccg gcccggatct cgcggggaat ggggctctcg 11400
gatgtagatc tgcgatccgc cgttgttggg ggagatgatg gggggtttaa aatttccgcc 11460
gtgctaaaca agatcaggaa gaggggaaaa gggcactatg gtttatattt ttatatattt 11520
ctgctgcttc gtcaggctta gatgtgctag atctttcttt cttctttttg tgggtagaat 11580
ttgaatccct cagcattgtt catcggtagt ttttcttttc atgatttgtg acaaatgcag 11640
cctcgtgcgg agcttttttg taggtagaag tgatcaacca tggcgcaagt tagcagaatc 11700
tgcaatggtg tgcagaaccc atctcttatc tccaatctct cgaaatccag tcaacgcaaa 11760
tctcccttat cggtttctct gaagacgcag cagcatccac gagcttatcc gatttcgtcg 11820
tcgtggggat tgaagaagag tgggatgacg ttaattggct ctgagcttcg tcctcttaag 11880
gtcatgtctt ctgtttccac ggcgtgcatg cttcacggtg caagcagccg gcccgcaacc 11940
gcccgcaaat cctctggcct ttccggaacc gtccgcattc ccggcgacaa gtcgatctcc 12000
caccggtcct tcatgttcgg cggtctcgcg agcggtgaaa cgcgcatcac cggccttctg 12060
gaaggcgagg acgtcatcaa tacgggcaag gccatgcagg cgatgggcgc ccgcatccgt 12120
aaggaaggcg acacctggat catcgatggc gtcggcaatg gcggcctcct ggcgcctgag 12180
gcgccgctcg atttcggcaa tgccgccacg ggctgccgcc tgacgatggg cctcgtcggg 12240
gtctacgatt tcgacagcac cttcatcggc gacgcctcgc tcacaaagcg cccgatgggc 12300
cgcgtgttga acccgctgcg cgaaatgggc gtgcaggtga aatcggaaga cggtgaccgt 12360
cttcccgtta ccttgcgcgg gccgaagacg ccgacgccga tcacctaccg cgtgccgatg 12420
gcctccgcac aggtgaagtc cgccgtgctg ctcgccggcc tcaacacgcc cggcatcacg 12480
acggtcatcg agccgatcat gacgcgcgat catacggaaa agatgctgca gggctttggc 12540
gccaacctta ccgtcgagac ggatgcggac ggcgtgcgca ccatccgcct ggaaggccgc 12600
ggcaagctca ccggccaagt catcgacgtg ccgggcgacc cgtcctcgac ggccttcccg 12660
ctggttgcgg ccctgcttgt tccgggctcc gacgtcacca tcctcaacgt gctgatgaac 12720
cccacccgca ccggcctcat cctgacgctg caggaaatgg gcgccgacat cgaagtcatc 12780
aacccgcgcc ttgccggcgg cgaagacgtg gcggacctgc gcgttcgctc ctccacgctg 12840
aagggcgtca cggtgccgga agaccgcgcg ccttcgatga tcgacgaata tccgattctc 12900
gctgtcgccg ccgccttcgc ggaaggggcg accgtgatga acggtctgga agaactccgc 12960
gtcaaggaaa gcgaccgcct ctcggccgtc gccaatggcc tcaagctcaa tggcgtggat 13020
tgcgatgagg gcgagacgtc gctcgtcgtg cgtggccgcc ctgacggcaa ggggctcggc 13080
aacgcctcgg gcgccgccgt cgccacccat ctcgatcacc gcatcgccat gagcttcctc 13140
gtcatgggcc tcgtgtcgga aaaccctgtc acggtggacg atgccacgat gatcgccacg 13200
agcttcccgg agttcatgga cctgatggcc gggctgggcg cgaagatcga actctccgat 13260
acgaaggctg cctgatgagc tcgaattccc gatcgttcaa acatttggca ataaagtttc 13320
ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac 13380
gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg 13440
attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac 13500
taggataaat tatcgcgcgc ggtgtcatct atgttactag atcggggata acgatcaagc 13560
tataacttcg tataatgtat gctatacgaa gttatc 13596
<210>214
<211>13559
<212>DNA
<213>人工序列
<220>
<223>pMON68066;图49;Zm-ASA2-CTP+18::AgroAS(S51C)-nno的表达载体
<400>214
gcgccaaatc gtgaagtttc tcatctaagc ccccatttgg acgtgaatgt agacacgtcg 60
aaataaagat ttccgaatta gaataatttg tttattgctt tcgcctataa atacgacgga 120
tcgtaatttg tcgttttatc aaaatgtact ttcattttat aataacgctg cggacatcta 180
catttttgaa ttgaaaaaaa attggtaatt actctttctt tttctccata ttgaccatca 240
tactcattgc tgatccatgt agatttcccg gacatgaagc catttacaat tgaatatatc 300
ctgccgccgc tgccgctttg cacccggtgg agcttgcatg ttggtttcta cgcagaactg 360
agccggttag gcagataatt tccattgaga actgagccat gtgcaccttc cccccaacac 420
ggtgagcgac ggggcaacgg agtgatccac atgggacttt tcctagcttg gctgccattt 480
ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt 540
tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg 600
cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa 660
aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg 720
cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc 780
tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac 840
cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag 900
gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc 960
ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct 1020
catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccggccgcg 1080
gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc 1140
cagcccagcg gcgagggcaa ccagcccggt gagcgtcgga aagggtcgat cgaccgatgc 1200
ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg 1260
ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct 1320
gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg 1380
cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac 1440
gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct 1500
tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg 1560
gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc 1620
agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc 1680
tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg 1740
taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg 1800
ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat 1860
tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat 1920
atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc 1980
ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg 2040
ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg 2100
ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta 2160
aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg 2220
atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc 2280
ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg 2340
ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt 2400
catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa 2460
acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct 2520
ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga 2580
ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct 2640
ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 2700
acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca 2760
gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta 2820
ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 2880
atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 2940
cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 3000
gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 3060
ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 3120
agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 3180
tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 3240
ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 3300
gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 3360
ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 3420
gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 3480
aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 3540
aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 3600
ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 3660
gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 3720
gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 3780
tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 3840
ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 3900
ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 3960
atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 4020
ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 4080
tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 4140
attgctgcag gtcgggagca caggatgacg cctaacaatt cattcaagcc gacaccgctt 4200
cgcggcgcgg cttaattcag gagttaaaca tcatgaggga agcggtgatc gccgaagtat 4260
cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg 4320
ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt 4380
tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc 4440
ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca 4500
ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg 4560
gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg 4620
atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg 4680
cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct 4740
taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt 4800
tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg 4860
actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctaggcagg 4920
cttatcttgg acaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgttc 4980
actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctaaca attcgttcaa 5040
gccgacgccg cttcgcggcg cggcttaact caagcgttag atgctgcagg catcgtggtg 5100
tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 5160
acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgaggat 5220
ttttcggcgc tgcgctacgt ccgcgaccgc gttgagggat caagccacag cagcccactc 5280
gaccttctag ccgacccaga cgagccaagg gatctttttg gaatgctgct ccgtcgtcag 5340
gctttccgac gtttgggtgg ttgaacagaa gtcattatcg cacggaatgc caagcactcc 5400
cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac 5460
gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa 5520
tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatcccca 5580
tcaagctagc ttctgcaggt cctgctcgag gtcactaagc aactaacttt gaggaatgag 5640
gtgatgatga attaactcac tccattccac aaaccaaaca aaaatttgag gagtgagaag 5700
atgattgact atctcattcc tcaaaccaaa cacctcaaat atatctgcta tcgggattgg 5760
cattcctgta tccctacgcc cgtgtacccc ctgtttagag aacctccaaa ggtataagat 5820
ggcgaagatt attgttgtct tgtctttcat catatatcga gtctttccct aggatattat 5880
tattggcaat gagcattaca cggttaatcg attgagagaa catgcatctc accttcagca 5940
aataattacg ataatccata ttttacgctt cgtaacttct catgagtttc gatatacaaa 6000
tttgttttct ggacacccta ccattcatcc tcttcggaga agagaggaag tgtcctcaat 6060
ttaaatatgt tgtcatgctg tagttcttca caaaatctca acaggtacca agcacattgt 6120
ttccacaaat tatattttag tcacaataaa tctatattat tattaatata ctaaaactat 6180
actgacgctc agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc 6240
agaaaatagt gagacacgga agacaaaaga agtaaaagag gcccggacta cggcccacat 6300
gagattcggc cccgccacct ccggcaacca gcggccgatc caacggcagt gcgcgcacac 6360
acacaacctc gtatatatcg ccgcgcggaa gcggcgcgac cgaggaagcc ttgtcctcga 6420
caccccctac acaggtgtcg cgctgccccc gacacgagtc ccgcatgcgt cccacgcggc 6480
cgcgccagat cccgcctccg cgcgttgcca cgccctctat aaacacccag ctctccctcg 6540
ccctcatcta cctcactcgt agtcgtagct caagcatcag cggcagcggc agcggcagga 6600
tctctgggca gcgtgcgcac gtggggtatc tagctcgctc tgctagccta ccaatcgaat 6660
tcctgcaggt cgactctaga ggatctaccg tcttcggtac gcgctcactc cgccctctgc 6720
ctttgttact gccacgtttc tctgaatgct ctcttgtgtg gtgattgctg agagtggttt 6780
agctggatct agaattacac tctgaaatcg tgttctgcct gtgctgatta cttgccgtcc 6840
tttgtagcag caaaatatag ggacatggta gtacgaaacg aagatagaac ctacacagca 6900
atacgagaaa tgtgtaattt ggtgcttagc ggtatttatt taagcacatg ttggtgttat 6960
agggcacttg gattcagaag tttgctgtta atttaggcac aggcttcata ctacatgggt 7020
caatagtata gggattcata ttataggcga tactataata atttgttcgt ctgcagagct 7080
tattatttgc caaaattaga tattcctatt ctgtttttgt ttgtgtgctg ttaaattgtt 7140
aacgcctgaa ggaataaata taaatgacga aattttgatg tttatctctg ctcctttatt 7200
gtgaccataa gtcaagatca gatgcacttg ttttaaatat tgttgtctga agaaataagt 7260
actgacagta ttttgatgca ttgatctgct tgtttgttgt aacaaaattt aaaaataaag 7320
agtttccttt ttgttgctct ccttacctcc tgatggtatc tagtatctac caactgacac 7380
tatattgctt ctctttacat acgtatcttg ctcgatgcct tctccctagt gttgaccagt 7440
gttactcaca tagtctttgc tcatttcatt gtaatgcaga taccaagcgg cctctagagg 7500
actccgatct atggaatccc tagccgccac ctccgtgttc gcgccctccc gcgtcgccgt 7560
cccggcggcg cgggccctgg ttagggcggg gacggtggta ccaaccaggc ggacgagcag 7620
ccggagcgga accagcgggg tgaaatgctc tgctgccgtg acgccgcagg cgagcccagt 7680
gattagcagg agcgctgcgg cggccatggt gaccatcatt caggacgacg gcgctgagac 7740
ctacgagact aagggcggta tccaagtgag ccgtaagcgt aggcccactg actacgctaa 7800
cgccatcgac aactacatcg agaagctaga ctcccatcgc ggcgctgtgt tctcctgcaa 7860
ctacgaatac cctgggcgct acacgaggtg ggataccgcc atcgtcgatc ctccattggg 7920
catctcctgt tttgggcgta agatgtggat cgaggcgtac aacggccgtg gcgaagtctt 7980
gctggacttc atcacggaga agctcaaggc cacaccggac ctcaccctcg gcgcttcctc 8040
gacccgccgc ctcgacctta cggtcaacga gccggaccgc gtgttcaccg aggaagagcg 8100
tagcaagatc ccgactgtct tcaccgcgct cagagccatc gtggacctat tctactcttc 8160
tgcggacagc gccatcgggt tgttcggtgc cttcggttac gacctcgcgt tccagttcga 8220
cgccatcaag ctctcgctcg cgcggccgga ggaccagcga gacatggtgc tcttcctccc 8280
tgacgagatc ctggtcgtcg atcactattc cgcgaaggcg tggatcgacc ggtacgactt 8340
cgagaaggat ggcatgacca cggatggcaa gagcagcgac atcactcccg acccattcaa 8400
gaccaccgac accatccctc caaagggcga tcaccgccct ggcgagtatt ccgaactcgt 8460
ggtgaaggcc aaggaatcct tccggcgcgg cgatctgttt gaggtggttc ccggccaaaa 8520
gttcatggag aggtgcgagt cgaatccgtc tgcgatcagt cgccgactga aagcgatcaa 8580
cccgagcccg tattccttct tcatcaacct cggcgatcag gaatatctgg tcggagcctc 8640
acccgagatg ttcgtcaggg tctccggccg ccggatcgag acgtgcccaa tttccggaac 8700
catcaagcgc ggagatgacc cgatagccga ctctgagcag atcctgaaac tcttgaacag 8760
caagaaggac gagtccgagc tgactatgtg ctcagatgtg gaccgaaacg acaagtcacg 8820
tgtctgcgag cccggtagcg tcaaggtcat tggccgccgt cagatcgaga tgtactccag 8880
gctgattcac acggtcgatc atatcgaagg gcggctgcgc gacgatatgg acgcattcga 8940
cgggttcctc agtcacgcct gggccgttac tgtcaccgga gcgcctaagc tctgggctat 9000
gaggttcatc gagggccacg agaagagccc tagggcttgg tatggtggtg ccatcggcat 9060
ggttgggttc aacggcgaca tgaacaccgg gctgacgctc cggaccatca ggatcaaaga 9120
cggcattgcc gaggtgaggg ccggtgccac gcttctcaac gatagcaacc ctcaggagga 9180
agaggcggag accgagctga aagcctctgc gatgatctcc gcgattagag atgcaaaggg 9240
tacgaacagt gctgccacca agcgggacgc agccaaggtg ggcaccggcg tcaagatttt 9300
acttgtcgat cacgaggact ccttcgtgca cactctggcg aactacttcc gccagacagg 9360
cgcgacggtc tccaccgtta ggtcaccggt ggccgctgac gtgttcgata ggttccagcc 9420
cgaccttgtg gtgctctctc ccggtcccgg ctcgcccacg gacttcgact gcaaggccac 9480
cattaaggcc gccagggcca gggatctgcc aatcttcggc gtttgcctcg ggcttcaggc 9540
attggccgag gcatacggtg gagagctgag gcagctcgcc gtcccgatgc acgggaagcc 9600
atcccgcatc agagtcctgg agcccggcct cgtcttctcc ggtctcggga aggaggtcac 9660
ggtcggtcgg tatcattcga tcttcgccga tccggcaacc ctcccgcgcg acttcatcat 9720
aaccgccgag tcggaggacg gaacgatcat gggaatcgaa cacgccaagg agcccgtagc 9780
tgcggttcag ttccaccctg agtccatcat gaccctcggt caagatgcgg gtatgcggat 9840
gatcgagaat gtggtggttc acctcacccg caaggccaag accaaggcag catgataggg 9900
atccgttggc aatgcggata aagaataact aaataaataa ataaataaat tgcaagcaat 9960
tgcgttgctg ctatgtactg taaaagtttc ttataatatc agttctgaat gctaaggaca 10020
tccctcaaga tggtctttct atttttgtgt tcccgttcca atgtactgtt ggtatcctct 10080
tggagattca tcaatatgag aaaacagaga atggacaacc ctcccttatc ttatggctcg 10140
agcggccgct ctagaactag tggatccccc ccttaattaa gggggctgca ggaattcata 10200
acttcgtata atgtatgcta tacgaagtta tgtttcgagg tcattcatat gcttgagaag 10260
agagtcggga tagtccaaaa taaaacaaag gtaagattac ctggtcaaaa gtgaaaacat 10320
cagttaaaag gtggtataaa gtaaaatatc ggtaataaaa ggtggcccaa agtgaaattt 10380
actcttttct actattataa aaattgagga tgtttttgtc ggtactttga tacgtcattt 10440
ttgtatgaat tggtttttaa gtttattcgc ttttggaaat gcatatctgt atttgagtcg 10500
ggttttaagt tcgtttgctt ttgtaaatac agagggattt gtataagaaa tatctttaga 10560
aaaacccata tgctaatttg acataatttt tgagaaaaat atatattcag gcgaattctc 10620
acaatgaaca ataataagat taaaatagct ttcccccgtt gcagcgcatg ggtatttttt 10680
ctagtaaaaa taaaagataa acttagactc aaaacattta caaaaacaac ccctaaagtt 10740
cctaaagccc aaagtgctat ccacgatcca tagcaagccc agcccaaccc aacccaaccc 10800
aacccacccc agtccagcca actggacaat agtctccaca cccccccact atcaccgtga 10860
gttgtccgca cgcaccgcac gtctcgcagc caaaaaaaaa aagaaagaaa aaaaagaaaa 10920
agaaaaaaca gcaggtgggt ccgggtcgtg ggggccggaa acgcgaggag gatcgcgagc 10980
cagcgacgag gccggccctc cctccgcttc caaagaaacg ccccccatcg ccactatata 11040
catacccccc cctctcctcc catcccccca accctaccac caccaccacc accacctcca 11100
cctcctcccc cctcgctgcc ggacgacgag ctcctccccc ctccccctcc gccgccgccg 11160
cgccggtaac caccccgccc ctctcctctt tctttctccg tttttttttc cgtctcggtc 11220
tcgatctttg gccttggtag tttgggtggg cgagaggcgg cttcgtgcgc gcccagatcg 11280
gtgcgcggga ggggcgggat ctcgcggctg gggctctcgc cggcgtggat ccggcccgga 11340
tctcgcgggg aatggggctc tcggatgtag atctgcgatc cgccgttgtt gggggagatg 11400
atggggggtt taaaatttcc gccgtgctaa acaagatcag gaagagggga aaagggcact 11460
atggtttata tttttatata tttctgctgc ttcgtcaggc ttagatgtgc tagatctttc 11520
tttcttcttt ttgtgggtag aatttgaatc cctcagcatt gttcatcggt agtttttctt 11580
ttcatgattt gtgacaaatg cagcctcgtg cggagctttt ttgtaggtag aagtgatcaa 11640
ccatggcgca agttagcaga atctgcaatg gtgtgcagaa cccatctctt atctccaatc 11700
tctcgaaatc cagtcaacgc aaatctccct tatcggtttc tctgaagacg cagcagcatc 11760
cacgagctta tccgatttcg tcgtcgtggg gattgaagaa gagtgggatg acgttaattg 11820
gctctgagct tcgtcctctt aaggtcatgt cttctgtttc cacggcgtgc atgcttcacg 11880
gtgcaagcag ccggcccgca accgcccgca aatcctctgg cctttccggaaccgtccgca 11940
ttcccggcga caagtcgatc tcccaccggt ccttcatgtt cggcggtctc gcgagcggtg 12000
aaacgcgcat caccggcctt ctggaaggcg aggacgtcat caatacgggc aaggccatgc 12060
aggcgatggg cgcccgcatc cgtaaggaag gcgacacctg gatcatcgat ggcgtcggca 12120
atggcggcct cctggcgcct gaggcgccgc tcgatttcgg caatgccgcc acgggctgcc 12180
gcctgacgat gggcctcgtc ggggtctacg atttcgacag caccttcatc ggcgacgcct 12240
cgctcacaaa gcgcccgatg ggccgcgtgt tgaacccgct gcgcgaaatg ggcgtgcagg 12300
tgaaatcgga agacggtgac cgtcttcccg ttaccttgcg cgggccgaag acgccgacgc 12360
cgatcaccta ccgcgtgccg atggcctccg cacaggtgaa gtccgccgtg ctgctcgccg 12420
gcctcaacac gcccggcatc acgacggtca tcgagccgat catgacgcgc gatcatacgg 12480
aaaagatgct gcagggcttt ggcgccaacc ttaccgtcga gacggatgcg gacggcgtgc 12540
gcaccatccg cctggaaggc cgcggcaagc tcaccggcca agtcatcgac gtgccgggcg 12600
acccgtcctc gacggccttc ccgctggttg cggccctgct tgttccgggc tccgacgtca 12660
ccatcctcaa cgtgctgatg aaccccaccc gcaccggcct catcctgacg ctgcaggaaa 12720
tgggcgccga catcgaagtc atcaacccgc gccttgccgg cggcgaagac gtggcggacc 12780
tgcgcgttcg ctcctccacg ctgaagggcg tcacggtgcc ggaagaccgc gcgccttcga 12840
tgatcgacga atatccgatt ctcgctgtcg ccgccgcctt cgcggaaggg gcgaccgtga 12900
tgaacggtct ggaagaactc cgcgtcaagg aaagcgaccg cctctcggcc gtcgccaatg 12960
gcctcaagct caatggcgtg gattgcgatg agggcgagac gtcgctcgtc gtgcgtggcc 13020
gccctgacgg caaggggctc ggcaacgcct cgggcgccgc cgtcgccacc catctcgatc 13080
accgcatcgc catgagcttc ctcgtcatgg gcctcgtgtc ggaaaaccct gtcacggtgg 13140
acgatgccac gatgatcgcc acgagcttcc cggagttcat ggacctgatg gccgggctgg 13200
gcgcgaagat cgaactctcc gatacgaagg ctgcctgatg agctcgaatt cccgatcgtt 13260
caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta 13320
tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt 13380
tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag 13440
aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac 13500
tagatcgggg ataacgatca agctataact tcgtataatg tatgctatac gaagttatc 13559
<210>215
<211>13905
<212>DNA
<213>人工序列
<220>
<223>pMON69757;图33;含有AgroAS(F298W)突变等位基因的构建体的的表达载体
<400>215
gtcctccgat cgaggatttt tcggcgctgc gctacgtccg cgaccgcgtt gagggatcaa 60
gccacagcag cccactcgac cttctagccg acccagacga gccaagggat ctttttggaa 120
tgctgctccg tcgtcaggct ttccgacgtt tgggtggttg aacagaagtc attatcgcac 180
ggaatgccaa gcactcccga ggggaaccct gtggttggca tgcacataca aatggacgaa 240
cggataaacc ttttcacgcc cttttaaata tccgattatt ctaataaacg ctcttttctc 300
ttaggtttac ccgccaatat atcctgtcaa acactgatag tttaaactga aggcgggaaa 360
cgacaatctg atccccatca agctagcttc tgcaggtcct gctcgaggtc actaagcaac 420
taactttgag gaatgaggtg atgatgaatt aactcactcc attccacaaa ccaaacaaaa 480
atttgaggag tgagaagatg attgactatc tcattcctca aaccaaacac ctcaaatata 540
tctgctatcg ggattggcat tcctgtatcc ctacgcccgt gtaccccctg tttagagaac 600
ctccaaaggt ataagatggc gaagattatt gttgtcttgt ctttcatcat atatcgagtc 660
tttccctagg atattattat tggcaatgag cattacacgg ttaatcgatt gagagaacat 720
gcatctcacc ttcagcaaat aattacgata atccatattt tacgcttcgt aacttctcat 780
gagtttcgat atacaaattt gttttctgga caccctacca ttcatcctct tcggagaaga 840
gaggaagtgt cctcaattta aatatgttgt catgctgtag ttcttcacaa aatctcaaca 900
ggtaccaagc acattgtttc cacaaattat attttagtca caataaatct atattattat 960
taatatacta aaactatact gacgctcaga tgcttttact agttcttgct agtatgtgat 1020
gtaggtctac gtggaccaga aaatagtgag acacggaaga caaaagaagt aaaagaggcc 1080
cggactacgg cccacatgag attcggcccc gccacctccg gcaaccagcg gccgatccaa 1140
cggcagtgcg cgcacacaca caacctcgta tatatcgccg cgcggaagcg gcgcgaccga 1200
ggaagccttg tcctcgacac cccctacaca ggtgtcgcgc tgcccccgac acgagtcccg 1260
catgcgtccc acgcggccgc gccagatccc gcctccgcgc gttgccacgc cctctataaa 1320
cacccagctc tccctcgccc tcatctacct cactcgtagt cgtagctcaa gcatcagcgg 1380
cagcggcagc ggcaggatct ctgggcagcg tgcgcacgtg gggtatctag ctcgctctgc 1440
tagcctacca atcgaattcc tgcaggtcga ctctagagga tctaccgtct tcggtacgcg 1500
ctcactccgc cctctgcctt tgttactgcc acgtttctct gaatgctctc ttgtgtggtg 1560
attgctgaga gtggtttagc tggatctaga attacactct gaaatcgtgt tctgcctgtg 1620
ctgattactt gccgtccttt gtagcagcaa aatataggga catggtagta cgaaacgaag 1680
atagaaccta cacagcaata cgagaaatgt gtaatttggt gcttagcggt atttatttaa 1740
gcacatgttg gtgttatagg gcacttggat tcagaagttt gctgttaatt taggcacagg 1800
cttcatacta catgggtcaa tagtataggg attcatatta taggcgatac tataataatt 1860
tgttcgtctg cagagcttat tatttgccaa aattagatat tcctattctg tttttgtttg 1920
tgtgctgtta aattgttaac gcctgaagga ataaatataa atgacgaaat tttgatgttt 1980
atctctgctc ctttattgtg accataagtc aagatcagat gcacttgttt taaatattgt 2040
tgtctgaaga aataagtact gacagtattt tgatgcattg atctgcttgt ttgttgtaac 2100
aaaatttaaa aataaagagt ttcctttttg ttgctctcct tacctcctga tggtatctag 2160
tatctaccaa ctgacactat attgcttctc tttacatacg tatcttgctc gatgccttct 2220
ccctagtgtt gaccagtgtt actcacatag tctttgctca tttcattgta atgcagatac 2280
caagcggcct ctagaggact ccgatctatg gaatccctag ccgccacctc cgtgttcgcg 2340
ccctcccgcg tcgccgtccc ggcggcgcgg gccctggtta gggcggggac ggtggtacca 2400
accaggcgga cgagcagccg gagcggaacc agcggggtga aatgctctgc tgccgtgacg 2460
ccgcaggcga gcccagtgat tagcaggagc gctgcggcgg ccatggtaac gatcattcag 2520
gatgacggag cggagaccta cgagacgaaa ggcggcatcc aggtcagccg aaagcgccgg 2580
cccaccgatt atgccaacgc catcgataat tacatcgaaa agcttgattc ccatcgcggc 2640
gcggtttttt cgtccaacta tgaatatccg ggccgttaca cccgctggga tacggccatc 2700
gtcgatccgc cgctcggcat ttcctgtttt ggccgcaaga tgtggatcga agcctataat 2760
ggccgcggcg aagtgctgct cgatttcatt acggaaaagc tgaaggcgac acccgatctc 2820
accctcggcg cttcctcgac ccgccggctc gatcttaccg tcaacgaacc ggaccgtgtc 2880
ttcaccgaag aagaacgctc gaaaatcccg acggtcttca ccgctctcag agccatcgtc 2940
gacctcttct attcgagcgc ggattcggcc atcggcctgt tcggtgcctt cggttacgat 3000
ctcgccttcc agttcgacgc gatcaagctt tcgctggcgc gtccggaaga ccagcgtgac 3060
atggtgctgt ttctgcccga tgaaatcctc gtcgttgatc actattccgc caaggcctgg 3120
atcgaccgtt acgatttcga gaaggacggc atgacgacgg acggcaaatc ctccgacatt 3180
acccccgatc ccttcaagac caccgatacc atcccgccca agggcgatca ccgtcccggc 3240
gaatattccg agcttgtggt gaaggccaag gaaagcttcc gccgcggcga cctgttcgag 3300
gtcgttcccg gccagaaatt catggagcgt tgcgaaagca atccgtcggc gatttcccgc 3360
cgcctgaagg cgatcaaccc gtcgccctat tcctggttca tcaatctcgg cgatcaggaa 3420
tatctggtcg gcgcctcgcc ggaaatgttc gtgcgcgtct ccggccgtcg catcgagacc 3480
tgcccgatat caggcaccat caagcgcggc gacgatccga ttgccgacag cgagcagatt 3540
ttgaaactgc tcaactcgaa aaaggacgaa tccgaactga ccatgtgctc ggacgtggac 3600
cgcaacgaca agagccgcgt ctgcgagccg ggttcggtga aggtcattgg ccgccgccag 3660
atcgagatgt attcacgcct catccacacc gtcgatcaca tcgaaggccg cctgcgcgac 3720
gatatggacg cctttgacgg tttcctcagc cacgcctggg ccgtcaccgt caccggtgca 3780
ccaaagctgt gggccatgcg cttcatcgaa ggtcatgaaa agagcccgcg cgcctggtat 3840
ggcggtgcga tcggcatggt cggcttcaac ggcgacatga ataccggcct gacgctgcgc 3900
accatccgga tcaaggacgg tattgccgaa gtgcgcgccg gcgcgaccct gctcaatgat 3960
tccaacccgc aggaagaaga agccgaaacc gaactgaagg cctccgccat gatatcagcc 4020
attcgtgacg caaaaggcac caactctgcc gccaccaagc gtgatgccgc caaagtcggc 4080
accggcgtca agatcctgct cgtcgaccac gaagacagct tcgtgcacac gctggcgaat 4140
tatttccgcc agacgggcgc gacggtctcg accgtcagat caccggtcgc agccgacgtg 4200
ttcgatcgct tccagccgga cctcgttgtc ctgtcgcccg gacccggcag cccgacggat 4260
ttcgactgca aggcaacgat caaggccgcc cgcgcccgcg atctgccgat cttcggcgtt 4320
tgcctcggtc tgcaggcatt ggcagaagcc tatggcggcg agctgcgcca gcttgctgtg 4380
cccatgcacg gcaagccttc gcgcatccgc gtgctggaac ccggcctcgt cttctccggt 4440
ctcggcaagg aagtcacggt cggtcgttac cattcgatct tcgccgatcc cgccaccctg 4500
ccgcgtgatt tcatcatcac cgcagaaagc gaggacggca cgatcatggg catcgaacac 4560
gccaaggaac cggtggccgc cgttcagttc cacccggaat cgatcatgac gctcggacag 4620
gacgcgggca tgcggatgat cgagaatgtc gtggtgcatc tgacccgcaa ggcgaagacc 4680
aaggccgcgt gatggcgctc gatgacacgg ttatatcact agtgcggcca tcggatccgg 4740
ggatcgatga gctaagctag ctatatcatc aatttatgta ttacacataa tatcgcactc 4800
agtctttcat ctacggcaat gtaccagctg atataatcag ttattgaaat atttctgaat 4860
ttaaacttgc atcaataaat ttatgttttt gcttggacta taatacctga cttgttattt 4920
tatcaataaa tatttaaact atatttcttt caagatatca ttctttacaa gtatacgtgt 4980
ttaaattgaa taccataaat ttttattttt caaatacatg taaaattatg aaatgggagt 5040
ggtggcgacc gagctcaagc acacttcaat tcctataacg gaccaaatcg caaaaattat 5100
aataacatat tatttcatcc tggattaaaa gaaagtcacc ggggattatt ttgtgacgcc 5160
gattacatac ggcgacaata aagacattgg aaatcgtagt acatattgga atacactgat 5220
tatattaatg atgaatacat actttaatat ccttacgtag gatcgatccg aatttcgacc 5280
tcgagcggcc gctctagaac tagtggatcc cccccttaat taagggggct gcaggaattc 5340
ataacttcgt ataatgtatg ctatacgaag ttatgtttcg aggtcattca tatgcttgag 5400
aagagagtcg ggatagtcca aaataaaaca aaggtaagat tacctggtca aaagtgaaaa 5460
catcagttaa aaggtggtat aaagtaaaat atcggtaata aaaggtggcc caaagtgaaa 5520
tttactcttt tctactatta taaaaattga ggatgttttt gtcggtactt tgatacgtca 5580
tttttgtatg aattggtttt taagtttatt cgcttttgga aatgcatatc tgtatttgag 5640
tcgggtttta agttcgtttg cttttgtaaa tacagaggga tttgtataag aaatatcttt 5700
agaaaaaccc atatgctaat ttgacataat ttttgagaaa aatatatatt caggcgaatt 5760
ctcacaatga acaataataa gattaaaata gctttccccc gttgcagcgc atgggtattt 5820
tttctagtaa aaataaaaga taaacttaga ctcaaaacat ttacaaaaac aacccctaaa 5880
gttcctaaag cccaaagtgc tatccacgat ccatagcaag cccagcccaa cccaacccaa 5940
cccaacccac cccagtccag ccaactggac aatagtctcc acaccccccc actatcaccg 6000
tgagttgtcc gcacgcaccg cacgtctcgc agccaaaaaa aaaaagaaag aaaaaaaaga 6060
aaaagaaaaa acagcaggtg ggtccgggtc gtgggggccg gaaacgcgag gaggatcgcg 6120
agccagcgac gaggccggcc ctccctccgc ttccaaagaa acgcccccca tcgccactat 6180
atacataccc ccccctctcc tcccatcccc ccaaccctac caccaccacc accaccacct 6240
ccacctcctc ccccctcgct gccggacgac gagctcctcc cccctccccc tccgccgccg 6300
ccgcgccggt aaccaccccg cccctctcct ctttctttct ccgttttttt ttccgtctcg 6360
gtctcgatct ttggccttgg tagtttgggt gggcgagagg cggcttcgtg cgcgcccaga 6420
tcggtgcgcg ggaggggcgg gatctcgcgg ctggggctct cgccggcgtg gatccggccc 6480
ggatctcgcg gggaatgggg ctctcggatg tagatctgcg atccgccgtt gttgggggag 6540
atgatggggg gtttaaaatt tccgccgtgc taaacaagat caggaagagg ggaaaagggc 6600
actatggttt atatttttat atatttctgc tgcttcgtca ggcttagatg tgctagatct 6660
ttctttcttc tttttgtggg tagaatttga atccctcagc attgttcatc ggtagttttt 6720
cttttcatga tttgtgacaa atgcagcctc gtgcggagct tttttgtagg tagaagtgat 6780
caaccatggc gcaagttagc agaatctgca atggtgtgca gaacccatct cttatctcca 6840
atctctcgaa atccagtcaa cgcaaatctc ccttatcggt ttctctgaag acgcagcagc 6900
atccacgagc ttatccgatt tcgtcgtcgt ggggattgaa gaagagtggg atgacgttaa 6960
ttggctctga gcttcgtcct cttaaggtca tgtcttctgt ttccacggcg tgcatgcttc 7020
acggtgcaag cagccggccc gcaaccgccc gcaaatcctc tggcctttcc ggaaccgtcc 7080
gcattcccgg cgacaagtcg atctcccacc ggtccttcat gttcggcggt ctcgcgagcg 7140
gtgaaacgcg catcaccggc cttctggaag gcgaggacgt catcaatacg ggcaaggcca 7200
tgcaggcgat gggcgcccgc atccgtaagg aaggcgacac ctggatcatc gatggcgtcg 7260
gcaatggcgg cctcctggcg cctgaggcgc cgctcgattt cggcaatgcc gccacgggct 7320
gccgcctgac gatgggcctc gtcggggtct acgatttcga cagcaccttc atcggcgacg 7380
cctcgctcac aaagcgcccg atgggccgcg tgttgaaccc gctgcgcgaa atgggcgtgc 7440
aggtgaaatc ggaagacggt gaccgtcttc ccgttacctt gcgcgggccg aagacgccga 7500
cgccgatcac ctaccgcgtg ccgatggcct ccgcacaggt gaagtccgcc gtgctgctcg 7560
ccggcctcaa cacgcccggc atcacgacgg tcatcgagcc gatcatgacg cgcgatcata 7620
cggaaaagat gctgcagggc tttggcgcca accttaccgt cgagacggat gcggacggcg 7680
tgcgcaccat ccgcctggaa ggccgcggca agctcaccgg ccaagtcatc gacgtgccgg 7740
gcgacccgtc ctcgacggcc ttcccgctgg ttgcggccct gcttgttccg ggctccgacg 7800
tcaccatcct caacgtgctg atgaacccca cccgcaccgg cctcatcctg acgctgcagg 7860
aaatgggcgc cgacatcgaa gtcatcaacc cgcgccttgc cggcggcgaa gacgtggcgg 7920
acctgcgcgt tcgctcctcc acgctgaagg gcgtcacggt gccggaagac cgcgcgcctt 7980
cgatgatcga cgaatatccg attctcgctg tcgccgccgc cttcgcggaa ggggcgaccg 8040
tgatgaacgg tctggaagaa ctccgcgtca aggaaagcga ccgcctctcg gccgtcgcca 8100
atggcctcaa gctcaatggc gtggattgcg atgagggcga gacgtcgctc gtcgtgcgtg 8160
gccgccctga cggcaagggg ctcggcaacg cctcgggcgc cgccgtcgcc acccatctcg 8220
atcaccgcat cgccatgagc ttcctcgtca tgggcctcgt gtcggaaaac cctgtcacgg 8280
tggacgatgc cacgatgatc gccacgagct tcccggagtt catggacctg atggccgggc 8340
tgggcgcgaa gatcgaactc tccgatacga aggctgcctg atgagctcga attcccgatc 8400
gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga 8460
ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga 8520
cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga 8580
tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt 8640
tactagatcg gggataacga tcaagctata acttcgtata atgtatgcta tacgaagtta 8700
tcgcgccaaa tcgtgaagtt tctcatctaa gcccccattt ggacgtgaat gtagacacgt 8760
cgaaataaag atttccgaat tagaataatt tgtttattgc tttcgcctat aaatacgacg 8820
gatcgtaatt tgtcgtttta tcaaaatgta ctttcatttt ataataacgc tgcggacatc 8880
tacatttttg aattgaaaaa aaattggtaa ttactctttc tttttctcca tattgaccat 8940
catactcatt gctgatccat gtagatttcc cggacatgaa gccatttaca attgaatata 9000
tcctgccgcc gctgccgctt tgcacccggt ggagcttgca tgttggtttc tacgcagaac 9060
tgagccggtt aggcagataa tttccattga gaactgagcc atgtgcacct tccccccaac 9120
acggtgagcg acggggcaac ggagtgatcc acatgggact tttcctagct tggctgccat 9180
ttttggggtg aggccgttcg cggccgaggg gcgcagcccc tggggggatg ggaggcccgc 9240
gttagcgggc cgggagggtt cgagaagggg gggcaccccc cttcggcgtg cgcggtcacg 9300
cgcacagggc gcagccctgg ttaaaaacaa ggtttataaa tattggttta aaagcaggtt 9360
aaaagacagg ttagcggtgg ccgaaaaacg ggcggaaacc cttgcaaatg ctggattttc 9420
tgcctgtgga cagcccctca aatgtcaata ggtgcgcccc tcatctgtca gcactctgcc 9480
cctcaagtgt caaggatcgc gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat 9540
accgcagggc acttatcccc aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc 9600
aggcgttttc gccgatttgc gaggctggcc agctccacgt cgccggccga aatcgagcct 9660
gcccctcatc tgtcaacgcc gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc 9720
ctcatctgtc agtgagggcc aagttttccg cgaggtatcc acaacgccgg cggccggccg 9780
cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc 9840
gccagcccag cggcgagggc aaccagcccg gtgagcgtcg gaaagggtcg atcgaccgat 9900
gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt 9960
cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct 10020
ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct 10080
tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa 10140
acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt 10200
cttgctggcg ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc 10260
cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca 10320
tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc 10380
gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat 10440
tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg 10500
ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga 10560
attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac 10620
atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg 10680
tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg 10740
ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc 10800
tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg 10860
taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca 10920
ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga 10980
ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa 11040
cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt 11100
ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc 11160
aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt 11220
ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac 11280
gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac 11340
ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 11400
agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc 11460
cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg 11520
tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 11580
gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 11640
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 11700
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 11760
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 11820
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 11880
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 11940
tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 12000
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 12060
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 12120
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 12180
tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 12240
tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 12300
ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 12360
aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 12420
aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 12480
aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 12540
gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 12600
gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 12660
caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 12720
ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 12780
attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 12840
ccattgctgc aggtcgggag cacaggatga cgcctaacaa ttcattcaag ccgacaccgc 12900
ttcgcggcgc ggcttaattc aggagttaaa catcatgagg gaagcggtga tcgccgaagt 12960
atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct 13020
ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga 13080
tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga 13140
ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac 13200
cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt 13260
tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat 13320
tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc 13380
ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac 13440
cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac 13500
gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc 13560
cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctaggca 13620
ggcttatctt ggacaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt 13680
tcactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taatgtctaa caattcgttc 13740
aagccgacgc cgcttcgcgg cgcggcttaa ctcaagcgtt agatgctgca ggcatcgtgg 13800
tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 13860
ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcg 13905
<210>216
<211>13905
<212>DNA
<213>人工序列
<220>
<223>pMON69770;图37;含有AgroAS(S51C)非最优化等位基因的构建体的表达载体
<400>216
tcgaggattt ttcggcgctg cgctacgtcc gcgaccgcgt tgagggatca agccacagca 60
gcccactcga ccttctagcc gacccagacg agccaaggga tctttttgga atgctgctcc 120
gtcgtcaggc tttccgacgt ttgggtggtt gaacagaagt cattatcgca cggaatgcca 180
agcactcccg aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac 240
cttttcacgc ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta 300
cccgccaata tatcctgtca aacactgata gtttaaactg aaggcgggaa acgacaatct 360
gatccccatc aagctagctt ctgcaggtcc tgctcgaggt cactaagcaa ctaactttga 420
ggaatgaggt gatgatgaat taactcactc cattccacaa accaaacaaa aatttgagga 480
gtgagaagat gattgactat ctcattcctc aaaccaaaca cctcaaatat atctgctatc 540
gggattggca ttcctgtatc cctacgcccg tgtaccccct gtttagagaa cctccaaagg 600
tataagatgg cgaagattat tgttgtcttg tctttcatca tatatcgagt ctttccctag 660
gatattatta ttggcaatga gcattacacg gttaatcgat tgagagaaca tgcatctcac 720
cttcagcaaa taattacgat aatccatatt ttacgcttcg taacttctca tgagtttcga 780
tatacaaatt tgttttctgg acaccctacc attcatcctc ttcggagaag agaggaagtg 840
tcctcaattt aaatatgttg tcatgctgta gttcttcaca aaatctcaac aggtaccaag 900
cacattgttt ccacaaatta tattttagtc acaataaatc tatattatta ttaatatact 960
aaaactatac tgacgctcag atgcttttac tagttcttgc tagtatgtga tgtaggtcta 1020
cgtggaccag aaaatagtga gacacggaag acaaaagaag taaaagaggc ccggactacg 1080
gcccacatga gattcggccc cgccacctcc ggcaaccagc ggccgatcca acggcagtgc 1140
gcgcacacac acaacctcgt atatatcgcc gcgcggaagc ggcgcgaccg aggaagcctt 1200
gtcctcgaca ccccctacac aggtgtcgcg ctgcccccga cacgagtccc gcatgcgtcc 1260
cacgcggccg cgccagatcc cgcctccgcg cgttgccacg ccctctataa acacccagct 1320
ctccctcgcc ctcatctacc tcactcgtag tcgtagctca agcatcagcg gcagcggcag 1380
cggcaggatc tctgggcagc gtgcgcacgt ggggtatcta gctcgctctg ctagcctacc 1440
aatcgaattc ctgcaggtcg actctagagg atctaccgtc ttcggtacgc gctcactccg 1500
ccctctgcct ttgttactgc cacgtttctc tgaatgctct cttgtgtggt gattgctgag 1560
agtggtttag ctggatctag aattacactc tgaaatcgtg ttctgcctgt gctgattact 1620
tgccgtcctt tgtagcagca aaatataggg acatggtagt acgaaacgaa gatagaacct 1680
acacagcaat acgagaaatg tgtaatttgg tgcttagcgg tatttattta agcacatgtt 1740
ggtgttatag ggcacttgga ttcagaagtt tgctgttaat ttaggcacag gcttcatact 1800
acatgggtca atagtatagg gattcatatt ataggcgata ctataataat ttgttcgtct 1860
gcagagctta ttatttgcca aaattagata ttcctattct gtttttgttt gtgtgctgtt 1920
aaattgttaa cgcctgaagg aataaatata aatgacgaaa ttttgatgtt tatctctgct 1980
cctttattgt gaccataagt caagatcaga tgcacttgtt ttaaatattg ttgtctgaag 2040
aaataagtac tgacagtatt ttgatgcatt gatctgcttg tttgttgtaa caaaatttaa 2100
aaataaagag tttccttttt gttgctctcc ttacctcctg atggtatcta gtatctacca 2160
actgacacta tattgcttct ctttacatac gtatcttgct cgatgccttc tccctagtgt 2220
tgaccagtgt tactcacata gtctttgctc atttcattgt aatgcagata ccaagcggcc 2280
tctagaggac tccgatctat ggaatcccta gccgccacct ccgtgttcgc gccctcccgc 2340
gtcgccgtcc cggcggcgcg ggccctggtt agggcgggga cggtggtacc aaccaggcgg 2400
acgagcagcc ggagcggaac cagcggggtg aaatgctctg ctgccgtgac gccgcaggcg 2460
agcccagtga ttagcaggag cgctgcggcg gccatggtaa cgatcattca ggatgacgga 2520
gcggagacct acgagacgaa aggcggcatc caggtcagcc gaaagcgccg gcccaccgat 2580
tatgccaacg ccatcgataa ttacatcgaa aagcttgatt cccatcgcgg cgcggttttt 2640
tcgtgcaact atgaatatcc gggccgttac acccgctggg atacggccat cgtcgatccg 2700
ccgctcggca tttcctgttt tggccgcaag atgtggatcg aagcctataa tggccgcggc 2760
gaagtgctgc tcgatttcat tacggaaaag ctgaaggcga cacccgatct caccctcggc 2820
gcttcctcga cccgccggct cgatcttacc gtcaacgaac cggaccgtgt cttcaccgaa 2880
gaagaacgct cgaaaatccc gacggtcttc accgctctca gagccatcgt cgacctcttc 2940
tattcgagcg cggattcggc catcggcctg ttcggtgcct tcggttacga tctcgccttc 3000
cagttcgacg cgatcaagct ttcgctggcg cgtccggaag accagcgtga catggtgctg 3060
tttctgcccg atgaaatcct cgtcgttgat cactattccg ccaaggcctg gatcgaccgt 3120
tacgatttcg agaaggacgg catgacgacg gacggcaaat cctccgacat tacccccgat 3180
cccttcaaga ccaccgatac catcccgccc aagggcgatc accgtcccgg cgaatattcc 3240
gagcttgtgg tgaaggccaa ggaaagcttc cgccgcggcg acctgttcga ggtcgttccc 3300
ggccagaaat tcatggagcg ttgcgaaagc aatccgtcgg cgatttcccg ccgcctgaag 3360
gcgatcaacc cgtcgcccta ttccttcttc atcaatctcg gcgatcagga atatctggtc 3420
ggcgcctcgc cggaaatgtt cgtgcgcgtc tccggccgtc gcatcgagac ctgcccgata 3480
tcaggcacca tcaagcgcgg cgacgatccg attgccgaca gcgagcagat tttgaaactg 3540
ctcaactcga aaaaggacga atccgaactg accatgtgct cggacgtgga ccgcaacgac 3600
aagagccgcg tctgcgagcc gggttcggtg aaggtcattg gccgccgcca gatcgagatg 3660
tattcacgcc tcatccacac cgtcgatcac atcgaaggcc gcctgcgcga cgatatggac 3720
gcctttgacg gtttcctcag ccacgcctgg gccgtcaccg tcaccggtgc accaaagctg 3780
tgggccatgc gcttcatcga aggtcatgaa aagagcccgc gcgcctggta tggcggtgcg 3840
atcggcatgg tcggcttcaa cggcgacatg aataccggcc tgacgctgcg caccatccgg 3900
atcaaggacg gtattgccga agtgcgcgcc ggcgcgaccc tgctcaatga ttccaacccg 3960
caggaagaag aagccgaaac cgaactgaag gcctccgcca tgatatcagc cattcgtgac 4020
gcaaaaggca ccaactctgc cgccaccaag cgtgatgccg ccaaagtcgg caccggcgtc 4080
aagatcctgc tcgtcgacca cgaagacagc ttcgtgcaca cgctggcgaa ttatttccgc 4140
cagacgggcg cgacggtctc gaccgtcaga tcaccggtcg cagccgacgt gttcgatcgc 4200
ttccagccgg acctcgttgt cctgtcgccc ggacccggca gcccgacgga tttcgactgc 4260
aaggcaacga tcaaggccgc ccgcgcccgc gatctgccga tcttcggcgt ttgcctcggt 4320
ctgcaggcat tggcagaagc ctatggcggc gagctgcgcc agcttgctgt gcccatgcac 4380
ggcaagcctt cgcgcatccg cgtgctggaa cccggcctcg tcttctccgg tctcggcaag 4440
gaagtcacgg tcggtcgtta ccattcgatc ttcgccgatc ccgccaccct gccgcgtgat 4500
ttcatcatca ccgcagaaag cgaggacggc acgatcatgg gcatcgaaca cgccaaggaa 4560
ccggtggccg ccgttcagtt ccacccggaa tcgatcatga cgctcggaca ggacgcgggc 4620
atgcggatga tcgagaatgt cgtggtgcat ctgacccgca aggcgaagac caaggccgcg 4680
tgatggcgct cgatgacacg gttatatcac tagtgcggcc atcggatccg gggatcgatg 4740
agctaagcta gctatatcat caatttatgt attacacata atatcgcact cagtctttca 4800
tctacggcaa tgtaccagct gatataatca gttattgaaa tatttctgaa tttaaacttg 4860
catcaataaa tttatgtttt tgcttggact ataatacctg acttgttatt ttatcaataa 4920
atatttaaac tatatttctt tcaagatatc attctttaca agtatacgtg tttaaattga 4980
ataccataaa tttttatttt tcaaatacat gtaaaattat gaaatgggag tggtggcgac 5040
cgagctcaag cacacttcaa ttcctataac ggaccaaatc gcaaaaatta taataacata 5100
ttatttcatc ctggattaaa agaaagtcac cggggattat tttgtgacgc cgattacata 5160
cggcgacaat aaagacattg gaaatcgtag tacatattgg aatacactga ttatattaat 5220
gatgaataca tactttaata tccttacgta ggatcgatcc gaatttcgac ctcgagcggc 5280
cgctctagaa ctagtggatc ccccccttaa ttaagggggc tgcaggaatt cataacttcg 5340
tataatgtat gctatacgaa gttatgtttc gaggtcattc atatgcttga gaagagagtc 5400
gggatagtcc aaaataaaac aaaggtaaga ttacctggtc aaaagtgaaa acatcagtta 5460
aaaggtggta taaagtaaaa tatcggtaat aaaaggtggc ccaaagtgaa atttactctt 5520
ttctactatt ataaaaattg aggatgtttt tgtcggtact ttgatacgtc atttttgtat 5580
gaattggttt ttaagtttat tcgcttttgg aaatgcatat ctgtatttga gtcgggtttt 5640
aagttcgttt gcttttgtaa atacagaggg atttgtataa gaaatatctt tagaaaaacc 5700
catatgctaa tttgacataa tttttgagaa aaatatatat tcaggcgaat tctcacaatg 5760
aacaataata agattaaaat agctttcccc cgttgcagcg catgggtatt ttttctagta 5820
aaaataaaag ataaacttag actcaaaaca tttacaaaaa caacccctaa agttcctaaa 5880
gcccaaagtg ctatccacga tccatagcaa gcccagccca acccaaccca acccaaccca 5940
ccccagtcca gccaactgga caatagtctc cacacccccc cactatcacc gtgagttgtc 6000
cgcacgcacc gcacgtctcg cagccaaaaa aaaaaagaaa gaaaaaaaag aaaaagaaaa 6060
aacagcaggt gggtccgggt cgtgggggcc ggaaacgcga ggaggatcgc gagccagcga 6120
cgaggccggc cctccctccg cttccaaaga aacgcccccc atcgccacta tatacatacc 6180
cccccctctc ctcccatccc cccaacccta ccaccaccac caccaccacc tccacctcct 6240
cccccctcgc tgccggacga cgagctcctc ccccctcccc ctccgccgcc gccgcgccgg 6300
taaccacccc gcccctctcc tctttctttc tccgtttttt tttccgtctc ggtctcgatc 6360
tttggccttg gtagtttggg tgggcgagag gcggcttcgt gcgcgcccag atcggtgcgc 6420
gggaggggcg ggatctcgcg gctggggctc tcgccggcgt ggatccggcc cggatctcgc 6480
ggggaatggg gctctcggat gtagatctgc gatccgccgt tgttggggga gatgatgggg 6540
ggtttaaaat ttccgccgtg ctaaacaaga tcaggaagag gggaaaaggg cactatggtt 6600
tatattttta tatatttctg ctgcttcgtc aggcttagat gtgctagatc tttctttctt 6660
ctttttgtgg gtagaatttg aatccctcag cattgttcat cggtagtttt tcttttcatg 6720
atttgtgaca aatgcagcct cgtgcggagc ttttttgtag gtagaagtga tcaaccatgg 6780
cgcaagttag cagaatctgc aatggtgtgc agaacccatc tcttatctcc aatctctcga 6840
aatccagtca acgcaaatct cccttatcgg tttctctgaa gacgcagcag catccacgag 6900
cttatccgat ttcgtcgtcg tggggattga agaagagtgg gatgacgtta attggctctg 6960
agcttcgtcc tcttaaggtc atgtcttctg tttccacggc gtgcatgctt cacggtgcaa 7020
gcagccggcc cgcaaccgcc cgcaaatcct ctggcctttc cggaaccgtc cgcattcccg 7080
gcgacaagtc gatctcccac cggtccttca tgttcggcgg tctcgcgagc ggtgaaacgc 7140
gcatcaccgg ccttctggaa ggcgaggacg tcatcaatac gggcaaggcc atgcaggcga 7200
tgggcgcccg catccgtaag gaaggcgaca cctggatcat cgatggcgtc ggcaatggcg 7260
gcctcctggc gcctgaggcg ccgctcgatt tcggcaatgc cgccacgggc tgccgcctga 7320
cgatgggcct cgtcggggtc tacgatttcg acagcacctt catcggcgac gcctcgctca 7380
caaagcgccc gatgggccgc gtgttgaacc cgctgcgcga aatgggcgtg caggtgaaat 7440
cggaagacgg tgaccgtctt cccgttacct tgcgcgggcc gaagacgccg acgccgatca 7500
cctaccgcgt gccgatggcc tccgcacagg tgaagtccgc cgtgctgctc gccggcctca 7560
acacgcccgg catcacgacg gtcatcgagc cgatcatgac gcgcgatcat acggaaaaga 7620
tgctgcaggg ctttggcgcc aaccttaccg tcgagacgga tgcggacggc gtgcgcacca 7680
tccgcctgga aggccgcggc aagctcaccg gccaagtcat cgacgtgccg ggcgacccgt 7740
cctcgacggc cttcccgctg gttgcggccc tgcttgttcc gggctccgac gtcaccatcc 7800
tcaacgtgct gatgaacccc acccgcaccg gcctcatcct gacgctgcag gaaatgggcg 7860
ccgacatcga agtcatcaac ccgcgccttg ccggcggcga agacgtggcg gacctgcgcg 7920
ttcgctcctc cacgctgaag ggcgtcacgg tgccggaaga ccgcgcgcct tcgatgatcg 7980
acgaatatcc gattctcgct gtcgccgccg ccttcgcgga aggggcgacc gtgatgaacg 8040
gtctggaaga actccgcgtc aaggaaagcg accgcctctc ggccgtcgcc aatggcctca 8100
agctcaatgg cgtggattgc gatgagggcg agacgtcgct cgtcgtgcgt ggccgccctg 8160
acggcaaggg gctcggcaac gcctcgggcg ccgccgtcgc cacccatctc gatcaccgca 8220
tcgccatgag cttcctcgtc atgggcctcg tgtcggaaaa ccctgtcacg gtggacgatg 8280
ccacgatgat cgccacgagc ttcccggagt tcatggacct gatggccggg ctgggcgcga 8340
agatcgaact ctccgatacg aaggctgcct gatgagctcg aattcccgat cgttcaaaca 8400
tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat 8460
aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta 8520
tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca 8580
aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc 8640
ggggataacg atcaagctat aacttcgtat aatgtatgct atacgaagtt atcgcgccaa 8700
atcgtgaagt ttctcatcta agcccccatt tggacgtgaa tgtagacacg tcgaaataaa 8760
gatttccgaa ttagaataat ttgtttattg ctttcgccta taaatacgac ggatcgtaat 8820
ttgtcgtttt atcaaaatgt actttcattt tataataacg ctgcggacat ctacattttt 8880
gaattgaaaa aaaattggta attactcttt ctttttctcc atattgacca tcatactcat 8940
tgctgatcca tgtagatttc ccggacatga agccatttac aattgaatat atcctgccgc 9000
cgctgccgct ttgcacccgg tggagcttgc atgttggttt ctacgcagaa ctgagccggt 9060
taggcagata atttccattg agaactgagc catgtgcacc ttccccccaa cacggtgagc 9120
gacggggcaa cggagtgatc cacatgggac ttttcctagc ttggctgcca tttttggggt 9180
gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg cgttagcggg 9240
ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg 9300
cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt taaaagacag 9360
gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg 9420
acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg 9480
tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg 9540
cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat caggcgtttt 9600
cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc tgcccctcat 9660
ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt 9720
cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccggcc gcggtgtctc 9780
gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca 9840
gcggcgaggg caaccagccc ggtgagcgtc ggaaagggtc gatcgaccga tgcccttgag 9900
agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact 9960
tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat 10020
tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt 10080
cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg 10140
cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc 10200
gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat 10260
cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca 10320
gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt 10380
cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc 10440
cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc 10500
gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc 10560
aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc 10620
gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca 10680
cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt 10740
actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa 10800
acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg 10860
gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc 10920
tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg 10980
atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt 11040
aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt 11100
atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa 11160
aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa 11220
ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct 11280
gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 11340
atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 11400
cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt 11460
agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag 11520
tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc 11580
gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 11640
tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 11700
agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 11760
cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 11820
ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 11880
tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 11940
gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 12000
gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 12060
gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 12120
ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 12180
ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 12240
ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 12300
gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 12360
ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 12420
tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 12480
ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 12540
gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 12600
tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 12660
cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 12720
ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 12780
gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg 12840
caggtcggga gcacaggatg acgcctaaca attcattcaa gccgacaccg cttcgcggcg 12900
cggcttaatt caggagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca 12960
actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca 13020
tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt 13080
tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga 13140
aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt 13200
gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg 13260
gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc 13320
tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga 13380
actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct 13440
atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg 13500
catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc 13560
aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctaggc aggcttatct 13620
tggacaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg ttcactacgt 13680
gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta acaattcgtt caagccgacg 13740
ccgcttcgcg gcgcggctta actcaagcgt tagatgctgc aggcatcgtg gtgtcacgct 13800
cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 13860
cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccga 13905
<210>217
<211>13905
<212>DNA
<213>人工序列
<220>
<223>pMON69768;图36;含有AgroAS(S51F)突变等位基因的构建体的表达载体
<400>217
tcgaggattt ttcggcgctg cgctacgtcc gcgaccgcgt tgagggatca agccacagca 60
gcccactcga ccttctagcc gacccagacg agccaaggga tctttttgga atgctgctcc 120
gtcgtcaggc tttccgacgt ttgggtggtt gaacagaagt cattatcgca cggaatgcca 180
agcactcccg aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac 240
cttttcacgc ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta 300
cccgccaata tatcctgtca aacactgata gtttaaactg aaggcgggaa acgacaatct 360
gatccccatc aagctagctt ctgcaggtcc tgctcgaggt cactaagcaa ctaactttga 420
ggaatgaggt gatgatgaat taactcactc cattccacaa accaaacaaa aatttgagga 480
gtgagaagat gattgactat ctcattcctc aaaccaaaca cctcaaatat atctgctatc 540
gggattggca ttcctgtatc cctacgcccg tgtaccccct gtttagagaa cctccaaagg 600
tataagatgg cgaagattat tgttgtcttg tctttcatca tatatcgagt ctttccctag 660
gatattatta ttggcaatga gcattacacg gttaatcgat tgagagaaca tgcatctcac 720
cttcagcaaa taattacgat aatccatatt ttacgcttcg taacttctca tgagtttcga 780
tatacaaatt tgttttctgg acaccctacc attcatcctc ttcggagaag agaggaagtg 840
tcctcaattt aaatatgttg tcatgctgta gttcttcaca aaatctcaac aggtaccaag 900
cacattgttt ccacaaatta tattttagtc acaataaatc tatattatta ttaatatact 960
aaaactatac tgacgctcag atgcttttac tagttcttgc tagtatgtga tgtaggtcta 1020
cgtggaccag aaaatagtga gacacggaag acaaaagaag taaaagaggc ccggactacg 1080
gcccacatga gattcggccc cgccacctcc ggcaaccagc ggccgatcca acggcagtgc 1140
gcgcacacac acaacctcgt atatatcgcc gcgcggaagc ggcgcgaccg aggaagcctt 1200
gtcctcgaca ccccctacac aggtgtcgcg ctgcccccga cacgagtccc gcatgcgtcc 1260
cacgcggccg cgccagatcc cgcctccgcg cgttgccacg ccctctataa acacccagct 1320
ctccctcgcc ctcatctacc tcactcgtag tcgtagctca agcatcagcg gcagcggcag 1380
cggcaggatc tctgggcagc gtgcgcacgt ggggtatcta gctcgctctg ctagcctacc 1440
aatcgaattc ctgcaggtcg actctagagg atctaccgtc ttcggtacgc gctcactccg 1500
ccctctgcct ttgttactgc cacgtttctc tgaatgctct cttgtgtggt gattgctgag 1560
agtggtttag ctggatctag aattacactc tgaaatcgtg ttctgcctgt gctgattact 1620
tgccgtcctt tgtagcagca aaatataggg acatggtagt acgaaacgaa gatagaacct 1680
acacagcaat acgagaaatg tgtaatttgg tgcttagcgg tatttattta agcacatgtt 1740
ggtgttatag ggcacttgga ttcagaagtt tgctgttaat ttaggcacag gcttcatact 1800
acatgggtca atagtatagg gattcatatt ataggcgata ctataataat ttgttcgtct 1860
gcagagctta ttatttgcca aaattagata ttcctattct gtttttgttt gtgtgctgtt 1920
aaattgttaa cgcctgaagg aataaatata aatgacgaaa ttttgatgtt tatctctgct 1980
cctttattgt gaccataagt caagatcaga tgcacttgtt ttaaatattg ttgtctgaag 2040
aaataagtac tgacagtatt ttgatgcatt gatctgcttg tttgttgtaa caaaatttaa 2100
aaataaagag tttccttttt gttgctctcc ttacctcctg atggtatcta gtatctacca 2160
actgacacta tattgcttct ctttacatac gtatcttgct cgatgccttc tccctagtgt 2220
tgaccagtgt tactcacata gtctttgctc atttcattgt aatgcagata ccaagcggcc 2280
tctagaggac tccgatctat ggaatcccta gccgccacct ccgtgttcgc gccctcccgc 2340
gtcgccgtcc cggcggcgcg ggccctggtt agggcgggga cggtggtacc aaccaggcgg 2400
acgagcagcc ggagcggaac cagcggggtg aaatgctctg ctgccgtgac gccgcaggcg 2460
agcccagtga ttagcaggag cgctgcggcg gccatggtaa cgatcattca ggatgacgga 2520
gcggagacct acgagacgaa aggcggcatc caggtcagcc gaaagcgccg gcccaccgat 2580
tatgccaacg ccatcgataa ttacatcgaa aagcttgatt cccatcgcgg cgcggttttt 2640
tcgttcaact atgaatatcc gggccgttac acccgctggg atacggccat cgtcgatccg 2700
ccgctcggca tttcctgttt tggccgcaag atgtggatcg aagcctataa tggccgcggc 2760
gaagtgctgc tcgatttcat tacggaaaag ctgaaggcga cacccgatct caccctcggc 2820
gcttcctcga cccgccggct cgatcttacc gtcaacgaac cggaccgtgt cttcaccgaa 2880
gaagaacgct cgaaaatccc gacggtcttc accgctctca gagccatcgt cgacctcttc 2940
tattcgagcg cggattcggc catcggcctg ttcggtgcct tcggttacga tctcgccttc 3000
cagttcgacg cgatcaagct ttcgctggcg cgtccggaag accagcgtga catggtgctg 3060
tttctgcccg atgaaatcct cgtcgttgat cactattccg ccaaggcctg gatcgaccgt 3120
tacgatttcg agaaggacgg catgacgacg gacggcaaat cctccgacat tacccccgat 3180
cccttcaaga ccaccgatac catcccgccc aagggcgatc accgtcccgg cgaatattcc 3240
gagcttgtgg tgaaggccaa ggaaagcttc cgccgcggcg acctgttcga ggtcgttccc 3300
ggccagaaat tcatggagcg ttgcgaaagc aatccgtcgg cgatttcccg ccgcctgaag 3360
gcgatcaacc cgtcgcccta ttccttcttc atcaatctcg gcgatcagga atatctggtc 3420
ggcgcctcgc cggaaatgtt cgtgcgcgtc tccggccgtc gcatcgagac ctgcccgata 3480
tcaggcacca tcaagcgcgg cgacgatccg attgccgaca gcgagcagat tttgaaactg 3540
ctcaactcga aaaaggacga atccgaactg accatgtgct cggacgtgga ccgcaacgac 3600
aagagccgcg tctgcgagcc gggttcggtg aaggtcattg gccgccgcca gatcgagatg 3660
tattcacgcc tcatccacac cgtcgatcac atcgaaggcc gcctgcgcga cgatatggac 3720
gcctttgacg gtttcctcag ccacgcctgg gccgtcaccg tcaccggtgc accaaagctg 3780
tgggccatgc gcttcatcga aggtcatgaa aagagcccgc gcgcctggta tggcggtgcg 3840
atcggcatgg tcggcttcaa cggcgacatg aataccggcc tgacgctgcg caccatccgg 3900
atcaaggacg gtattgccga agtgcgcgcc ggcgcgaccc tgctcaatga ttccaacccg 3960
caggaagaag aagccgaaac cgaactgaag gcctccgcca tgatatcagc cattcgtgac 4020
gcaaaaggca ccaactctgc cgccaccaag cgtgatgccg ccaaagtcgg caccggcgtc 4080
aagatcctgc tcgtcgacca cgaagacagc ttcgtgcaca cgctggcgaa ttatttccgc 4140
cagacgggcg cgacggtctc gaccgtcaga tcaccggtcg cagccgacgt gttcgatcgc 4200
ttccagccgg acctcgttgt cctgtcgccc ggacccggca gcccgacgga tttcgactgc 4260
aaggcaacga tcaaggccgc ccgcgcccgc gatctgccga tcttcggcgt ttgcctcggt 4320
ctgcaggcat tggcagaagc ctatggcggc gagctgcgcc agcttgctgt gcccatgcac 4380
ggcaagcctt cgcgcatccg cgtgctggaa cccggcctcg tcttctccgg tctcggcaag 4440
gaagtcacgg tcggtcgtta ccattcgatc ttcgccgatc ccgccaccct gccgcgtgat 4500
ttcatcatca ccgcagaaag cgaggacggc acgatcatgg gcatcgaaca cgccaaggaa 4560
ccggtggccg ccgttcagtt ccacccggaa tcgatcatga cgctcggaca ggacgcgggc 4620
atgcggatga tcgagaatgt cgtggtgcat ctgacccgca aggcgaagac caaggccgcg 4680
tgatggcgct cgatgacacg gttatatcac tagtgcggcc atcggatccg gggatcgatg 4740
agctaagcta gctatatcat caatttatgt attacacata atatcgcact cagtctttca 4800
tctacggcaa tgtaccagct gatataatca gttattgaaa tatttctgaa tttaaacttg 4860
catcaataaa tttatgtttt tgcttggact ataatacctg acttgttatt ttatcaataa 4920
atatttaaac tatatttctt tcaagatatc attctttaca agtatacgtg tttaaattga 4980
ataccataaa tttttatttt tcaaatacat gtaaaattat gaaatgggag tggtggcgac 5040
cgagctcaag cacacttcaa ttcctataac ggaccaaatc gcaaaaatta taataacata 5100
ttatttcatc ctggattaaa agaaagtcac cggggattat tttgtgacgc cgattacata 5160
cggcgacaat aaagacattg gaaatcgtag tacatattgg aatacactga ttatattaat 5220
gatgaataca tactttaata tccttacgta ggatcgatcc gaatttcgac ctcgagcggc 5280
cgctctagaa ctagtggatc ccccccttaa ttaagggggc tgcaggaatt cataacttcg 5340
tataatgtat gctatacgaa gttatgtttc gaggtcattc atatgcttga gaagagagtc 5400
gggatagtcc aaaataaaac aaaggtaaga ttacctggtc aaaagtgaaa acatcagtta 5460
aaaggtggta taaagtaaaa tatcggtaat aaaaggtggc ccaaagtgaa atttactctt 5520
ttctactatt ataaaaattg aggatgtttt tgtcggtact ttgatacgtc atttttgtat 5580
gaattggttt ttaagtttat tcgcttttgg aaatgcatat ctgtatttga gtcgggtttt 5640
aagttcgttt gcttttgtaa atacagaggg atttgtataa gaaatatctt tagaaaaacc 5700
catatgctaa tttgacataa tttttgagaa aaatatatat tcaggcgaat tctcacaatg 5760
aacaataata agattaaaat agctttcccc cgttgcagcg catgggtatt ttttctagta 5820
aaaataaaag ataaacttag actcaaaaca tttacaaaaa caacccctaa agttcctaaa 5880
gcccaaagtg ctatccacga tccatagcaa gcccagccca acccaaccca acccaaccca 5940
ccccagtcca gccaactgga caatagtctc cacacccccc cactatcacc gtgagttgtc 6000
cgcacgcacc gcacgtctcg cagccaaaaa aaaaaagaaa gaaaaaaaag aaaaagaaaa 6060
aacagcaggt gggtccgggt cgtgggggcc ggaaacgcga ggaggatcgc gagccagcga 6120
cgaggccggc cctccctccg cttccaaaga aacgcccccc atcgccacta tatacatacc 6180
cccccctctc ctcccatccc cccaacccta ccaccaccac caccaccacc tccacctcct 6240
cccccctcgc tgccggacga cgagctcctc ccccctcccc ctccgccgcc gccgcgccgg 6300
taaccacccc gcccctctcc tctttctttc tccgtttttt tttccgtctc ggtctcgatc 6360
tttggccttg gtagtttggg tgggcgagag gcggcttcgt gcgcgcccag atcggtgcgc 6420
gggaggggcg ggatctcgcg gctggggctc tcgccggcgt ggatccggcc cggatctcgc 6480
ggggaatggg gctctcggat gtagatctgc gatccgccgt tgttggggga gatgatgggg 6540
ggtttaaaat ttccgccgtg ctaaacaaga tcaggaagag gggaaaaggg cactatggtt 6600
tatattttta tatatttctg ctgcttcgtc aggcttagat gtgctagatc tttctttctt 6660
ctttttgtgg gtagaatttg aatccctcag cattgttcat cggtagtttt tcttttcatg 6720
atttgtgaca aatgcagcct cgtgcggagc ttttttgtag gtagaagtga tcaaccatgg 6780
cgcaagttag cagaatctgc aatggtgtgc agaacccatc tcttatctcc aatctctcga 6840
aatccagtca acgcaaatct cccttatcgg tttctctgaa gacgcagcag catccacgag 6900
cttatccgat ttcgtcgtcg tggggattga agaagagtgg gatgacgtta attggctctg 6960
agcttcgtcc tcttaaggtc atgtcttctg tttccacggc gtgcatgctt cacggtgcaa 7020
gcagccggcc cgcaaccgcc cgcaaatcct ctggcctttc cggaaccgtc cgcattcccg 7080
gcgacaagtc gatctcccac cggtccttca tgttcggcgg tctcgcgagc ggtgaaacgc 7140
gcatcaccgg ccttctggaa ggcgaggacg tcatcaatac gggcaaggcc atgcaggcga 7200
tgggcgcccg catccgtaag gaaggcgaca cctggatcat cgatggcgtc ggcaatggcg 7260
gcctcctggc gcctgaggcg ccgctcgatt tcggcaatgc cgccacgggc tgccgcctga 7320
cgatgggcct cgtcggggtc tacgatttcg acagcacctt catcggcgac gcctcgctca 7380
caaagcgccc gatgggccgc gtgttgaacc cgctgcgcga aatgggcgtg caggtgaaat 7440
cggaagacgg tgaccgtctt cccgttacct tgcgcgggcc gaagacgccg acgccgatca 7500
cctaccgcgt gccgatggcc tccgcacagg tgaagtccgc cgtgctgctc gccggcctca 7560
acacgcccgg catcacgacg gtcatcgagc cgatcatgac gcgcgatcat acggaaaaga 7620
tgctgcaggg ctttggcgcc aaccttaccg tcgagacgga tgcggacggc gtgcgcacca 7680
tccgcctgga aggccgcggc aagctcaccg gccaagtcat cgacgtgccg ggcgacccgt 7740
cctcgacggc cttcccgctg gttgcggccc tgcttgttcc gggctccgac gtcaccatcc 7800
tcaacgtgct gatgaacccc acccgcaccg gcctcatcct gacgctgcag gaaatgggcg 7860
ccgacatcga agtcatcaac ccgcgccttg ccggcggcga agacgtggcg gacctgcgcg 7920
ttcgctcctc cacgctgaag ggcgtcacgg tgccggaaga ccgcgcgcct tcgatgatcg 7980
acgaatatcc gattctcgct gtcgccgccg ccttcgcgga aggggcgacc gtgatgaacg 8040
gtctggaaga actccgcgtc aaggaaagcg accgcctctc ggccgtcgcc aatggcctca 8100
agctcaatgg cgtggattgc gatgagggcg agacgtcgct cgtcgtgcgt ggccgccctg 8160
acggcaaggg gctcggcaac gcctcgggcg ccgccgtcgc cacccatctc gatcaccgca 8220
tcgccatgag cttcctcgtc atgggcctcg tgtcggaaaa ccctgtcacg gtggacgatg 8280
ccacgatgat cgccacgagc ttcccggagt tcatggacct gatggccggg ctgggcgcga 8340
agatcgaact ctccgatacg aaggctgcct gatgagctcg aattcccgat cgttcaaaca 8400
tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat 8460
aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta 8520
tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca 8580
aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc 8640
ggggataacg atcaagctat aacttcgtat aatgtatgct atacgaagtt atcgcgccaa 8700
atcgtgaagt ttctcatcta agcccccatt tggacgtgaa tgtagacacg tcgaaataaa 8760
gatttccgaa ttagaataat ttgtttattg ctttcgccta taaatacgac ggatcgtaat 8820
ttgtcgtttt atcaaaatgt actttcattt tataataacg ctgcggacat ctacattttt 8880
gaattgaaaa aaaattggta attactcttt ctttttctcc atattgacca tcatactcat 8940
tgctgatcca tgtagatttc ccggacatga agccatttac aattgaatat atcctgccgc 9000
cgctgccgct ttgcacccgg tggagcttgc atgttggttt ctacgcagaa ctgagccggt 9060
taggcagata atttccattg agaactgagc catgtgcacc ttccccccaa cacggtgagc 9120
gacggggcaa cggagtgatc cacatgggac ttttcctagc ttggctgcca tttttggggt 9180
gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg cgttagcggg 9240
ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg 9300
cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt taaaagacag 9360
gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg 9420
acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg 9480
tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg 9540
cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat caggcgtttt 9600
cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc tgcccctcat 9660
ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt 9720
cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccggcc gcggtgtctc 9780
gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca 9840
gcggcgaggg caaccagccc ggtgagcgtc ggaaagggtc gatcgaccga tgcccttgag 9900
agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact 9960
tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat 10020
tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt 10080
cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg 10140
cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc 10200
gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat 10260
cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca 10320
gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt 10380
cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc 10440
cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc 10500
gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc 10560
aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc 10620
gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca 10680
cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt 10740
actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa 10800
acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg 10860
gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc 10920
tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg 10980
atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt 11040
aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt 11100
atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa 11160
aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa 11220
ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct 11280
gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 11340
atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 11400
cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt 11460
agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag 11520
tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc 11580
gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 11640
tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 11700
agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 11760
cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 11820
ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 11880
tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 11940
gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 12000
gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 12060
gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 12120
ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 12180
ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 12240
ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 12300
gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 12360
ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 12420
tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 12480
ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 12540
gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 12600
tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 12660
cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 12720
ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 12780
gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg 12840
caggtcggga gcacaggatg acgcctaaca attcattcaa gccgacaccg cttcgcggcg 12900
cggcttaatt caggagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca 12960
actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca 13020
tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt 13080
tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga 13140
aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt 13200
gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg 13260
gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc 13320
tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga 13380
actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct 13440
atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg 13500
catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc 13560
aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctaggc aggcttatct 13620
tggacaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg ttcactacgt 13680
gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta acaattcgtt caagccgacg 13740
ccgcttcgcg gcgcggctta actcaagcgt tagatgctgc aggcatcgtg gtgtcacgct 13800
cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 13860
cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccga 13905
<210>218
<211>13888
<212>DNA
<213>人工序列
<220>
<223>pMON78850;图47;含有苜蓿根瘤菌(Rhizobium meliloti)邻氨基苯甲酸合酶
野生型等位基因的构建体的表达载体
<400>218
ccaaatcgtg aagtttctca tctaagcccc catttggacg tgaatgtaga cacgtcgaaa 60
taaagatttc cgaattagaa taatttgttt attgctttcg cctataaata cgacggatcg 120
taatttgtcg ttttatcaaa atgtactttc attttataat aacgctgcgg acatctacat 180
ttttgaattg aaaaaaaatt ggtaattact ctttcttttt ctccatattg accatcatac 240
tcattgctga tccatgtaga tttcccggac atgaagccat ttacaattga atatatcctg 300
ccgccgctgc cgctttgcac ccggtggagc ttgcatgttg gtttctacgc agaactgagc 360
cggttaggca gataatttcc attgagaact gagccatgtg caccttcccc ccaacacggt 420
gagcgacggg gcaacggagt gatccacatg ggacttttcc tagcttggct gccatttttg 480
gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag 540
cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac 600
agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag 660
acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct 720
gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca 780
agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc 840
agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg 900
ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc 960
tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat 1020
ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc ggccgcggtg 1080
tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag 1140
cccagcggcg agggcaacca gcccggtgag cgtcggaaag ggtcgatcga ccgatgccct 1200
tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg 1260
cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg 1320
tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg 1380
tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt 1440
tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc 1500
tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg 1560
gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg 1620
gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga 1680
tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag 1740
gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca 1800
cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg 1860
agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc 1920
catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg 1980
gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg 2040
ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg 2100
caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag 2160
tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg 2220
ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg 2280
agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc 2340
cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat 2400
cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca 2460
ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga 2520
gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca 2580
cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg 2640
acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 2700
agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc 2760
acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg 2820
agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 2880
aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 2940
gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 3000
ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 3060
ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 3120
cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 3180
ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 3240
tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 3300
gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 3360
tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 3420
gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 3480
tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 3540
ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 3600
agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 3660
gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 3720
attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 3780
agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 3840
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 3900
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 3960
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 4020
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 4080
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 4140
gctgcaggtc gggagcacag gatgacgcct aacaattcat tcaagccgac accgcttcgc 4200
ggcgcggctt aattcaggag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga 4260
ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg 4320
tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc 4380
tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt 4440
tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg 4500
ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag 4560
aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc 4620
tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg 4680
aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa 4740
cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt 4800
cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact 4860
gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct aggcaggctt 4920
atcttggaca agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgttcact 4980
acgtgaaagg cgagatcacc aaggtagtcg gcaaataatg tctaacaatt cgttcaagcc 5040
gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg ctgcaggcat cgtggtgtca 5100
cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5160
tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgaggatttt 5220
tcggcgctgc gctacgtccg cgaccgcgtt gagggatcaa gccacagcag cccactcgac 5280
cttctagccg acccagacga gccaagggat ctttttggaa tgctgctccg tcgtcaggct 5340
ttccgacgtt tgggtggttg aacagaagtc attatcgcac ggaatgccaa gcactcccga 5400
ggggaaccct gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc 5460
cttttaaata tccgattatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat 5520
atcctgtcaa acactgatag tttaaactga aggcgggaaa cgacaatctg atccccatca 5580
agctagcttc tgcaggtcct gctcgaggtc actaagcaac taactttgag gaatgaggtg 5640
atgatgaatt aactcactcc attccacaaa ccaaacaaaa atttgaggag tgagaagatg 5700
attgactatc tcattcctca aaccaaacac ctcaaatata tctgctatcg ggattggcat 5760
tcctgtatcc ctacgcccgt gtaccccctg tttagagaac ctccaaaggt ataagatggc 5820
gaagattatt gttgtcttgt ctttcatcat atatcgagtc tttccctagg atattattat 5880
tggcaatgag cattacacgg ttaatcgatt gagagaacat gcatctcacc ttcagcaaat 5940
aattacgata atccatattt tacgcttcgt aacttctcat gagtttcgat atacaaattt 6000
gttttctgga caccctacca ttcatcctct tcggagaaga gaggaagtgt cctcaattta 6060
aatatgttgt catgctgtag ttcttcacaa aatctcaaca ggtaccaagc acattgtttc 6120
cacaaattat attttagtca caataaatct atattattat taatatacta aaactatact 6180
gacgctcaga tgcttttact agttcttgct agtatgtgat gtaggtctac gtggaccaga 6240
aaatagtgag acacggaaga caaaagaagt aaaagaggcc cggactacgg cccacatgag 6300
attcggcccc gccacctccg gcaaccagcg gccgatccaa cggcagtgcg cgcacacaca 6360
caacctcgta tatatcgccg cgcggaagcg gcgcgaccga ggaagccttg tcctcgacac 6420
cccctacaca ggtgtcgcgc tgcccccgac acgagtcccg catgcgtccc acgcggccgc 6480
gccagatccc gcctccgcgc gttgccacgc cctctataaa cacccagctc tccctcgccc 6540
tcatctacct cactcgtagt cgtagctcaa gcatcagcgg cagcggcagc ggcaggatct 6600
ctgggcagcg tgcgcacgtg gggtatctag ctcgctctgc tagcctacca atcgaattcc 6660
tgcaggtcga ctctagagga tctaccgtct tcggtacgcg ctcactccgc cctctgcctt 6720
tgttactgcc acgtttctct gaatgctctc ttgtgtggtg attgctgaga gtggtttagc 6780
tggatctaga attacactct gaaatcgtgt tctgcctgtg ctgattactt gccgtccttt 6840
gtagcagcaa aatataggga catggtagta cgaaacgaag atagaaccta cacagcaata 6900
cgagaaatgt gtaatttggt gcttagcggt atttatttaa gcacatgttg gtgttatagg 6960
gcacttggat tcagaagttt gctgttaatt taggcacagg cttcatacta catgggtcaa 7020
tagtataggg attcatatta taggcgatac tataataatt tgttcgtctg cagagcttat 7080
tatttgccaa aattagatat tcctattctg tttttgtttg tgtgctgtta aattgttaac 7140
gcctgaagga ataaatataa atgacgaaat tttgatgttt atctctgctc ctttattgtg 7200
accataagtc aagatcagat gcacttgttt taaatattgt tgtctgaaga aataagtact 7260
gacagtattt tgatgcattg atctgcttgt ttgttgtaac aaaatttaaa aataaagagt 7320
ttcctttttg ttgctctcct tacctcctga tggtatctag tatctaccaa ctgacactat 7380
attgcttctc tttacatacg tatcttgctc gatgccttct ccctagtgtt gaccagtgtt 7440
actcacatag tctttgctca tttcattgta atgcagatac caagcggcct ctagaggatc 7500
tccctagagg atctccagat ctatggaatc cctagccgcc acctccgtgt tcgcgccctc 7560
ccgcgtcgcc gtcccggcgg cgcgggccct ggttagggcg gggacggtgg taccaaccag 7620
gcggacgagc agccggagcg gaaccagcgg ggtgaaatgc tctgctgccg tgacgccgca 7680
ggcgagccca gtgattagca ggagcgctgc ggcggccatg gcagcggtaa ttctggaaga 7740
cggcgcggag agttatacca cgaagggtgg catcgtcgtc acccgcaggc ggcgtgaggc 7800
atcctacagc gacgcgatcg ccggttatgt cgaccggctg gacgaacgcc gcggcgcggt 7860
cttttcctcg aactacgaat atcccggccg ctatacccgc tgggacactg cggtggtcga 7920
cccgccgctt gccatctcct ccttcggtcg ctcgctctgg atcgaagcct ataacgaacg 7980
cggcgaagtg ctgctggcgc tgatcgccga ggatctgaag tccgttgccg acatcacgct 8040
cggctcactt gccgcccgcc gcctcgacct caccatcaac gagcccgatc gtgtcttcac 8100
cgaggaagag cggtcgaaga tgccgacggt ctttacggtt cttcgcgcgg tgacgaacct 8160
cttccactcg gaggaggact cgaacctcgg cctctatggc gccttcggct acgacctcgc 8220
cttccagttc gatgcgatcg aactgaagct ttcgcgtccg gacgaccagc gcgacatggt 8280
tctctttctg ccggacgaga tccttgtggt cgatcactat gcggccaagg cctggatcga 8340
ccgctacgat ttcgccaggg agaacctttc gaccgagggc aaggcagcgg acattgctcc 8400
cgagccgttc cgcagcgtcg acagcatccc gccgcacggg gatcaccgcc cgggcgaata 8460
tgccgagctc gtcgtcaagg cgaaggaaag cttccgtcgc ggcgatcttt tcgaagtggt 8520
gccggggcag aaattctacg agcgctgcga aagccgcccg tccgagattt ccaaccggct 8580
gaaggcgatc aatccgtcgc cctattcctt cttcatcaat ctcggcaacc aggaatatct 8640
cgtcggtgct tcgccggaga tgttcgtgcg cgtttccggc cggcgcatcg agacctgccc 8700
gatctccggt acgatcaagc gcggcgacga tccgatcgcc gacagcgagc agatcctgaa 8760
gctcttgaac tcgaagaagg acgagtccga gctcaccatg tgctcggacg tcgaccgcaa 8820
cgacaagagc cgggtctgcg tgccgggctc ggtcaaggtg atcggccggc gtcagatcga 8880
gatgtattcg cggctgatcc acacggtcga tcacatcgag gggcgcctgc gcgacgatat 8940
ggacgccttc gacgggttcc tcagccacgc ctgggcggtg accgttaccg gcgcgccaaa 9000
gctctgggcc atgcgcttca tcgagagcca cgagaagagc ccgcgtgcct ggtatggcgg 9060
cgcgatcggc atggtcggct tcaacggcga catgaatacc gggctgacct tgcgtaccat 9120
ccgcatcaag gacgggatcg ccgaggtgag ggcgggtgcg acgctcctct atgattccaa 9180
tccggaagaa gaagaagccg aaaccgaact gaaggcctct gccatgattg cagccatccg 9240
cgacgcgaaa tccgcaaaca gcgccaaatc cgcgcgcgat gtcgccgccg tcggcgccgg 9300
agtcagcatc ctgctcgtcg atcacgagga cagcttcgtc cataccctcg cgaactactt 9360
ccgccagacc ggcgcgtccg tcaccaccgt gcgcacgccg gtggccgagg aaatcttcga 9420
ccgggtcaag ccggacctcg tcgtgctttc gcccggtccc ggcaccccga aggacttcga 9480
ctgcaaggcg acgatcaaga aggcgcgggc gcgggacctg ccgatcttcg gcgtctgcct 9540
ggggctgcag gcgctcgcgg aggcctatgg cggcgacctt cgtcaactgg cgatcccgat 9600
gcatgggaag ccctcgcgca tccgcgtgct cgaacccggc atcgtcttct ccggcctcgg 9660
caaggaggtg acggtcgggc gctatcattc gattttcgcc gatccgtcca acctgccgcg 9720
cgaattcgtg atcacggccg aaagcgaaga tggtacgatc atgggcatcg aacacagcaa 9780
ggagccggtg gcggccgtgc agttccatcc ggaatcgatc atgacgctgg gcggcgacgc 9840
cggcatgcgg atgatcgaga acgtggttgc ccatctcgcc aagcgggcga agaccaaggc 9900
agcctgaact agatcggatc cggggatcga tgagctaagc tagctatatc atcaatttat 9960
gtattacaca taatatcgca ctcagtcttt catctacggc aatgtaccag ctgatataat 10020
cagttattga aatatttctg aatttaaact tgcatcaata aatttatgtt tttgcttgga 10080
ctataatacc tgacttgtta ttttatcaat aaatatttaa actatatttc tttcaagata 10140
tcattcttta caagtatacg tgtttaaatt gaataccata aatttttatt tttcaaatac 10200
atgtaaaatt atgaaatggg agtggtggcg accgagctca agcacacttc aattcctata 10260
acggaccaaa tcgcaaaaat tataataaca tattatttca tcctggatta aaagaaagtc 10320
accggggatt attttgtgac gccgattaca tacggcgaca ataaagacat tggaaatcgt 10380
agtacatatt ggaatacact gattatatta atgatgaata catactttaa tatccttacg 10440
taggatcgat ccgaatttcg acctcgagcg gccgctctag aactagtgga tccccccctt 10500
aattaagggg gctgcaggaa ttcataactt cgtataatgt atgctatacg aagttatgtt 10560
tcgaggtcat tcatatgctt gagaagagag tcgggatagt ccaaaataaa acaaaggtaa 10620
gattacctgg tcaaaagtga aaacatcagt taaaaggtgg tataaagtaa aatatcggta 10680
ataaaaggtg gcccaaagtg aaatttactc ttttctacta ttataaaaat tgaggatgtt 10740
tttgtcggta ctttgatacg tcatttttgt atgaattggt ttttaagttt attcgctttt 10800
ggaaatgcat atctgtattt gagtcgggtt ttaagttcgt ttgcttttgt aaatacagag 10860
ggatttgtat aagaaatatc tttagaaaaa cccatatgct aatttgacat aatttttgag 10920
aaaaatatat attcaggcga attctcacaa tgaacaataa taagattaaa atagctttcc 10980
cccgttgcag cgcatgggta ttttttctag taaaaataaa agataaactt agactcaaaa 11040
catttacaaa aacaacccct aaagttccta aagcccaaag tgctatccac gatccatagc 11100
aagcccagcc caacccaacc caacccaacc caccccagtc cagccaactg gacaatagtc 11160
tccacacccc cccactatca ccgtgagttg tccgcacgca ccgcacgtct cgcagccaaa 11220
aaaaaaaaga aagaaaaaaa agaaaaagaa aaaacagcag gtgggtccgg gtcgtggggg 11280
ccggaaacgc gaggaggatc gcgagccagc gacgaggccg gccctccctc cgcttccaaa 11340
gaaacgcccc ccatcgccac tatatacata cccccccctc tcctcccatc cccccaaccc 11400
taccaccacc accaccacca cctccacctc ctcccccctc gctgccggac gacgagctcc 11460
tcccccctcc ccctccgccg ccgccgcgcc ggtaaccacc ccgcccctct cctctttctt 11520
tctccgtttt tttttccgtc tcggtctcga tctttggcct tggtagtttg ggtgggcgag 11580
aggcggcttc gtgcgcgccc agatcggtgc gcgggagggg cgggatctcg cggctggggc 11640
tctcgccggc gtggatccgg cccggatctc gcggggaatg gggctctcgg atgtagatct 11700
gcgatccgcc gttgttgggg gagatgatgg ggggtttaaa atttccgccg tgctaaacaa 11760
gatcaggaag aggggaaaag ggcactatgg tttatatttt tatatatttc tgctgcttcg 11820
tcaggcttag atgtgctaga tctttctttc ttctttttgt gggtagaatt tgaatccctc 11880
agcattgttc atcggtagtt tttcttttca tgatttgtga caaatgcagc ctcgtgcgga 11940
gcttttttgt aggtagaagt gatcaaccat ggcgcaagtt agcagaatct gcaatggtgt 12000
gcagaaccca tctcttatct ccaatctctc gaaatccagt caacgcaaat ctcccttatc 12060
ggtttctctg aagacgcagc agcatccacg agcttatccg atttcgtcgt cgtggggatt 12120
gaagaagagt gggatgacgt taattggctc tgagcttcgt cctcttaagg tcatgtcttc 12180
tgtttccacg gcgtgcatgc ttcacggtgc aagcagccgg cccgcaaccg cccgcaaatc 12240
ctctggcctt tccggaaccg tccgcattcc cggcgacaag tcgatctccc accggtcctt 12300
catgttcggc ggtctcgcga gcggtgaaac gcgcatcacc ggccttctgg aaggcgagga 12360
cgtcatcaat acgggcaagg ccatgcaggc gatgggcgcc cgcatccgta aggaaggcga 12420
cacctggatc atcgatggcg tcggcaatgg cggcctcctg gcgcctgagg cgccgctcga 12480
tttcggcaat gccgccacgg gctgccgcct gacgatgggc ctcgtcgggg tctacgattt 12540
cgacagcacc ttcatcggcg acgcctcgct cacaaagcgc ccgatgggcc gcgtgttgaa 12600
cccgctgcgc gaaatgggcg tgcaggtgaa atcggaagac ggtgaccgtc ttcccgttac 12660
cttgcgcggg ccgaagacgc cgacgccgat cacctaccgc gtgccgatgg cctccgcaca 12720
ggtgaagtcc gccgtgctgc tcgccggcct caacacgccc ggcatcacga cggtcatcga 12780
gccgatcatg acgcgcgatc atacggaaaa gatgctgcag ggctttggcg ccaaccttac 12840
cgtcgagacg gatgcggacg gcgtgcgcac catccgcctg gaaggccgcg gcaagctcac 12900
cggccaagtc atcgacgtgc cgggcgaccc gtcctcgacg gccttcccgc tggttgcggc 12960
cctgcttgtt ccgggctccg acgtcaccat cctcaacgtg ctgatgaacc ccacccgcac 13020
cggcctcatc ctgacgctgc aggaaatggg cgccgacatc gaagtcatca acccgcgcct 13080
tgccggcggc gaagacgtgg cggacctgcg cgttcgctcc tccacgctga agggcgtcac 13140
ggtgccggaa gaccgcgcgc cttcgatgat cgacgaatat ccgattctcg ctgtcgccgc 13200
cgccttcgcg gaaggggcga ccgtgatgaa cggtctggaa gaactccgcg tcaaggaaag 13260
cgaccgcctc tcggccgtcg ccaatggcct caagctcaat ggcgtggatt gcgatgaggg 13320
cgagacgtcg ctcgtcgtgc gtggccgccc tgacggcaag gggctcggca acgcctcggg 13380
cgccgccgtc gccacccatc tcgatcaccg catcgccatg agcttcctcg tcatgggcct 13440
cgtgtcggaa aaccctgtca cggtggacga tgccacgatg atcgccacga gcttcccgga 13500
gttcatggac ctgatggccg ggctgggcgc gaagatcgaa ctctccgata cgaaggctgc 13560
ctgatgagct cgaattcccg atcgttcaaa catttggcaa taaagtttct taagattgaa 13620
tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt 13680
aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc 13740
gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt 13800
atcgcgcgcg gtgtcatcta tgttactaga tcggggataa cgatcaagct ataacttcgt 13860
ataatgtatg ctatacgaag ttatcgcg 13888
<210>219
<211>13888
<212>DNA
<213>人工序列
<220>
<223>pMON78851;图48;含有苜蓿根瘤菌(Rhizobium meliloti)邻氨基苯甲酸合酶
S51C等位基因的构建体的表达载体
<400>219
ccaaatcgtg aagtttctca tctaagcccc catttggacg tgaatgtaga cacgtcgaaa 60
taaagatttc cgaattagaa taatttgttt attgctttcg cctataaata cgacggatcg 120
taatttgtcg ttttatcaaa atgtactttc attttataat aacgctgcgg acatctacat 180
ttttgaattg aaaaaaaatt ggtaattact ctttcttttt ctccatattg accatcatac 240
tcattgctga tccatgtaga tttcccggac atgaagccat ttacaattga atatatcctg 300
ccgccgctgc cgctttgcac ccggtggagc ttgcatgttg gtttctacgc agaactgagc 360
cggttaggca gataatttcc attgagaact gagccatgtg caccttcccc ccaacacggt 420
gagcgacggg gcaacggagt gatccacatg ggacttttcc tagcttggct gccatttttg 480
gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag 540
cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac 600
agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag 660
acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct 720
gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca 780
agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc 840
agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg 900
ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc 960
tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat 1020
ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc ggccgcggtg 1080
tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag 1140
cccagcggcg agggcaacca gcccggtgag cgtcggaaag ggtcgatcga ccgatgccct 1200
tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg 1260
cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg 1320
tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg 1380
tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt 1440
tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc 1500
tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg 1560
gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg 1620
gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga 1680
tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag 1740
gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca 1800
cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg 1860
agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc 1920
catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg 1980
gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg 2040
ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg 2100
caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag 2160
tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg 2220
ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg 2280
agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc 2340
cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat 2400
cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca 2460
ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga 2520
gaaactcaac gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca 2580
cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg 2640
acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 2700
agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc 2760
acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg 2820
agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 2880
aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 2940
gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 3000
ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 3060
ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 3120
cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 3180
ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 3240
tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 3300
gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 3360
tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 3420
gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 3480
tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 3540
ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 3600
agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 3660
gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 3720
attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 3780
agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 3840
atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 3900
cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 3960
ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 4020
agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 4080
tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 4140
gctgcaggtc gggagcacag gatgacgcct aacaattcat tcaagccgac accgcttcgc 4200
ggcgcggctt aattcaggag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga 4260
ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg 4320
tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc 4380
tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt 4440
tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg 4500
ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag 4560
aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc 4620
tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg 4680
aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa 4740
cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt 4800
cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact 4860
gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct aggcaggctt 4920
atcttggaca agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgttcact 4980
acgtgaaagg cgagatcacc aaggtagtcg gcaaataatg tctaacaatt cgttcaagcc 5040
gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg ctgcaggcat cgtggtgtca 5100
cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5160
tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgaggatttt 5220
tcggcgctgc gctacgtccg cgaccgcgtt gagggatcaa gccacagcag cccactcgac 5280
cttctagccg acccagacga gccaagggat ctttttggaa tgctgctccg tcgtcaggct 5340
ttccgacgtt tgggtggttg aacagaagtc attatcgcac ggaatgccaa gcactcccga 5400
ggggaaccct gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc 5460
cttttaaata tccgattatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat 5520
atcctgtcaa acactgatag tttaaactga aggcgggaaa cgacaatctg atccccatca 5580
agctagcttc tgcaggtcct gctcgaggtc actaagcaac taactttgag gaatgaggtg 5640
atgatgaatt aactcactcc attccacaaa ccaaacaaaa atttgaggag tgagaagatg 5700
attgactatc tcattcctca aaccaaacac ctcaaatata tctgctatcg ggattggcat 5760
tcctgtatcc ctacgcccgt gtaccccctg tttagagaac ctccaaaggt ataagatggc 5820
gaagattatt gttgtcttgt ctttcatcat atatcgagtc tttccctagg atattattat 5880
tggcaatgag cattacacgg ttaatcgatt gagagaacat gcatctcacc ttcagcaaat 5940
aattacgata atccatattt tacgcttcgt aacttctcat gagtttcgat atacaaattt 6000
gttttctgga caccctacca ttcatcctct tcggagaaga gaggaagtgt cctcaattta 6060
aatatgttgt catgctgtag ttcttcacaa aatctcaaca ggtaccaagc acattgtttc 6120
cacaaattat attttagtca caataaatct atattattat taatatacta aaactatact 6180
gacgctcaga tgcttttact agttcttgct agtatgtgat gtaggtctac gtggaccaga 6240
aaatagtgag acacggaaga caaaagaagt aaaagaggcc cggactacgg cccacatgag 6300
attcggcccc gccacctccg gcaaccagcg gccgatccaa cggcagtgcg cgcacacaca 6360
caacctcgta tatatcgccg cgcggaagcg gcgcgaccga ggaagccttg tcctcgacac 6420
cccctacaca ggtgtcgcgc tgcccccgac acgagtcccg catgcgtccc acgcggccgc 6480
gccagatccc gcctccgcgc gttgccacgc cctctataaa cacccagctc tccctcgccc 6540
tcatctacct cactcgtagt cgtagctcaa gcatcagcgg cagcggcagc ggcaggatct 6600
ctgggcagcg tgcgcacgtg gggtatctag ctcgctctgc tagcctacca atcgaattcc 6660
tgcaggtcga ctctagagga tctaccgtct tcggtacgcg ctcactccgc cctctgcctt 6720
tgttactgcc acgtttctct gaatgctctc ttgtgtggtg attgctgaga gtggtttagc 6780
tggatctaga attacactct gaaatcgtgt tctgcctgtg ctgattactt gccgtccttt 6840
gtagcagcaa aatataggga catggtagta cgaaacgaag atagaaccta cacagcaata 6900
cgagaaatgt gtaatttggt gcttagcggt atttatttaa gcacatgttg gtgttatagg 6960
gcacttggat tcagaagttt gctgttaatt taggcacagg cttcatacta catgggtcaa 7020
tagtataggg attcatatta taggcgatac tataataatt tgttcgtctg cagagcttat 7080
tatttgccaa aattagatat tcctattctg tttttgtttg tgtgctgtta aattgttaac 7140
gcctgaagga ataaatataa atgacgaaat tttgatgttt atctctgctc ctttattgtg 7200
accataagtc aagatcagat gcacttgttt taaatattgt tgtctgaaga aataagtact 7260
gacagtattt tgatgcattg atctgcttgt ttgttgtaac aaaatttaaa aataaagagt 7320
ttcctttttg ttgctctcct tacctcctga tggtatctag tatctaccaa ctgacactat 7380
attgcttctc tttacatacg tatcttgctc gatgccttct ccctagtgtt gaccagtgtt 7440
actcacatag tctttgctca tttcattgta atgcagatac caagcggcct ctagaggatc 7500
tccctagagg atctccagat ctatggaatc cctagccgcc acctccgtgt tcgcgccctc 7560
ccgcgtcgcc gtcccggcgg cgcgggccct ggttagggcg gggacggtgg taccaaccag 7620
gcggacgagc agccggagcg gaaccagcgg ggtgaaatgc tctgctgccg tgacgccgca 7680
ggcgagccca gtgattagca ggagcgctgc ggcggccatg gcagcggtaa ttctggaaga 7740
cggcgcggag agttatacca cgaagggtgg catcgtcgtc acccgcaggc ggcgtgaggc 7800
atcctacagc gacgcgatcg ccggttatgt cgaccggctg gacgaacgcc gcggcgcggt 7860
cttttcctgc aactacgaat atcccggccg ctatacccgc tgggacactg cggtggtcga 7920
cccgccgctt gccatctcct ccttcggtcg ctcgctctgg atcgaagcct ataacgaacg 7980
cggcgaagtg ctgctggcgc tgatcgccga ggatctgaag tccgttgccg acatcacgct 8040
cggctcactt gccgcccgcc gcctcgacct caccatcaac gagcccgatc gtgtcttcac 8100
cgaggaagag cggtcgaaga tgccgacggt ctttacggtt cttcgcgcgg tgacgaacct 8160
cttccactcg gaggaggact cgaacctcgg cctctatggc gccttcggct acgacctcgc 8220
cttccagttc gatgcgatcg aactgaagct ttcgcgtccg gacgaccagc gcgacatggt 8280
tctctttctg ccggacgaga tccttgtggt cgatcactat gcggccaagg cctggatcga 8340
ccgctacgat ttcgccaggg agaacctttc gaccgagggc aaggcagcgg acattgctcc 8400
cgagccgttc cgcagcgtcg acagcatccc gccgcacggg gatcaccgcc cgggcgaata 8460
tgccgagctc gtcgtcaagg cgaaggaaag cttccgtcgc ggcgatcttt tcgaagtggt 8520
gccggggcag aaattctacg agcgctgcga aagccgcccg tccgagattt ccaaccggct 8580
gaaggcgatc aatccgtcgc cctattcctt cttcatcaat ctcggcaacc aggaatatct 8640
cgtcggtgct tcgccggaga tgttcgtgcg cgtttccggc cggcgcatcg agacctgccc 8700
gatctccggt acgatcaagc gcggcgacga tccgatcgcc gacagcgagc agatcctgaa 8760
gctcttgaac tcgaagaagg acgagtccga gctcaccatg tgctcggacg tcgaccgcaa 8820
cgacaagagc cgggtctgcg tgccgggctc ggtcaaggtg atcggccggc gtcagatcga 8880
gatgtattcg cggctgatcc acacggtcga tcacatcgag gggcgcctgc gcgacgatat 8940
ggacgccttc gacgggttcc tcagccacgc ctgggcggtg accgttaccg gcgcgccaaa 9000
gctctgggcc atgcgcttca tcgagagcca cgagaagagc ccgcgtgcct ggtatggcgg 9060
cgcgatcggc atggtcggct tcaacggcga catgaatacc gggctgacct tgcgtaccat 9120
ccgcatcaag gacgggatcg ccgaggtgag ggcgggtgcg acgctcctct atgattccaa 9180
tccggaagaa gaagaagccg aaaccgaact gaaggcctct gccatgattg cagccatccg 9240
cgacgcgaaa tccgcaaaca gcgccaaatc cgcgcgcgat gtcgccgccg tcggcgccgg 9300
agtcagcatc ctgctcgtcg atcacgagga cagcttcgtc cataccctcg cgaactactt 9360
ccgccagacc ggcgcgtccg tcaccaccgt gcgcacgccg gtggccgagg aaatcttcga 9420
ccgggtcaag ccggacctcg tcgtgctttc gcccggtccc ggcaccccga aggacttcga 9480
ctgcaaggcg acgatcaaga aggcgcgggc gcgggacctg ccgatcttcg gcgtctgcct 9540
ggggctgcag gcgctcgcgg aggcctatgg cggcgacctt cgtcaactgg cgatcccgat 9600
gcatgggaag ccctcgcgca tccgcgtgct cgaacccggc atcgtcttct ccggcctcgg 9660
caaggaggtg acggtcgggc gctatcattc gattttcgcc gatccgtcca acctgccgcg 9720
cgaattcgtg atcacggccg aaagcgaaga tggtacgatc atgggcatcg aacacagcaa 9780
ggagccggtg gcggccgtgc agttccatcc ggaatcgatc atgacgctgg gcggcgacgc 9840
cggcatgcgg atgatcgaga acgtggttgc ccatctcgcc aagcgggcga agaccaaggc 9900
agcctgaact agatcggatc cggggatcga tgagctaagc tagctatatc atcaatttat 9960
gtattacaca taatatcgca ctcagtcttt catctacggc aatgtaccag ctgatataat 10020
cagttattga aatatttctg aatttaaact tgcatcaata aatttatgtt tttgcttgga 10080
ctataatacc tgacttgtta ttttatcaat aaatatttaa actatatttc tttcaagata 10140
tcattcttta caagtatacg tgtttaaatt gaataccata aatttttatt tttcaaatac 10200
atgtaaaatt atgaaatggg agtggtggcg accgagctca agcacacttc aattcctata 10260
acggaccaaa tcgcaaaaat tataataaca tattatttca tcctggatta aaagaaagtc 10320
accggggatt attttgtgac gccgattaca tacggcgaca ataaagacat tggaaatcgt 10380
agtacatatt ggaatacact gattatatta atgatgaata catactttaa tatccttacg 10440
taggatcgat ccgaatttcg acctcgagcg gccgctctag aactagtgga tccccccctt 10500
aattaagggg gctgcaggaa ttcataactt cgtataatgt atgctatacg aagttatgtt 10560
tcgaggtcat tcatatgctt gagaagagag tcgggatagt ccaaaataaa acaaaggtaa 10620
gattacctgg tcaaaagtga aaacatcagt taaaaggtgg tataaagtaa aatatcggta 10680
ataaaaggtg gcccaaagtg aaatttactc ttttctacta ttataaaaat tgaggatgtt 10740
tttgtcggta ctttgatacg tcatttttgt atgaattggt ttttaagttt attcgctttt 10800
ggaaatgcat atctgtattt gagtcgggtt ttaagttcgt ttgcttttgt aaatacagag 10860
ggatttgtat aagaaatatc tttagaaaaa cccatatgct aatttgacat aatttttgag 10920
aaaaatatat attcaggcga attctcacaa tgaacaataa taagattaaa atagctttcc 10980
cccgttgcag cgcatgggta ttttttctag taaaaataaa agataaactt agactcaaaa 11040
catttacaaa aacaacccct aaagttccta aagcccaaag tgctatccac gatccatagc 11100
aagcccagcc caacccaacc caacccaacc caccccagtc cagccaactg gacaatagtc 11160
tccacacccc cccactatca ccgtgagttg tccgcacgca ccgcacgtct cgcagccaaa 11220
aaaaaaaaga aagaaaaaaa agaaaaagaa aaaacagcag gtgggtccgg gtcgtggggg 11280
ccggaaacgc gaggaggatc gcgagccagc gacgaggccg gccctccctc cgcttccaaa 11340
gaaacgcccc ccatcgccac tatatacata cccccccctc tcctcccatc cccccaaccc 11400
taccaccacc accaccacca cctccacctc ctcccccctc gctgccggac gacgagctcc 11460
tcccccctcc ccctccgccg ccgccgcgcc ggtaaccacc ccgcccctct cctctttctt 11520
tctccgtttt tttttccgtc tcggtctcga tctttggcct tggtagtttg ggtgggcgag 11580
aggcggcttc gtgcgcgccc agatcggtgc gcgggagggg cgggatctcg cggctggggc 11640
tctcgccggc gtggatccgg cccggatctc gcggggaatg gggctctcgg atgtagatct 11700
gcgatccgcc gttgttgggg gagatgatgg ggggtttaaa atttccgccg tgctaaacaa 11760
gatcaggaag aggggaaaag ggcactatgg tttatatttt tatatatttc tgctgcttcg 11820
tcaggcttag atgtgctaga tctttctttc ttctttttgt gggtagaatt tgaatccctc 11880
agcattgttc atcggtagtt tttcttttca tgatttgtga caaatgcagc ctcgtgcgga 11940
gcttttttgt aggtagaagt gatcaaccat ggcgcaagtt agcagaatct gcaatggtgt 12000
gcagaaccca tctcttatct ccaatctctc gaaatccagt caacgcaaat ctcccttatc 12060
ggtttctctg aagacgcagc agcatccacg agcttatccg atttcgtcgt cgtggggatt 12120
gaagaagagt gggatgacgt taattggctc tgagcttcgt cctcttaagg tcatgtcttc 12180
tgtttccacg gcgtgcatgc ttcacggtgc aagcagccgg cccgcaaccg cccgcaaatc 12240
ctctggcctt tccggaaccg tccgcattcc cggcgacaag tcgatctccc accggtcctt 12300
catgttcggc ggtctcgcga gcggtgaaac gcgcatcacc ggccttctgg aaggcgagga 12360
cgtcatcaat acgggcaagg ccatgcaggc gatgggcgcc cgcatccgta aggaaggcga 12420
cacctggatc atcgatggcg tcggcaatgg cggcctcctg gcgcctgagg cgccgctcga 12480
tttcggcaat gccgccacgg gctgccgcct gacgatgggc ctcgtcgggg tctacgattt 12540
cgacagcacc ttcatcggcg acgcctcgct cacaaagcgc ccgatgggcc gcgtgttgaa 12600
cccgctgcgc gaaatgggcg tgcaggtgaa atcggaagac ggtgaccgtc ttcccgttac 12660
cttgcgcggg ccgaagacgc cgacgccgat cacctaccgc gtgccgatgg cctccgcaca 12720
ggtgaagtcc gccgtgctgc tcgccggcct caacacgccc ggcatcacga cggtcatcga 12780
gccgatcatg acgcgcgatc atacggaaaa gatgctgcag ggctttggcg ccaaccttac 12840
cgtcgagacg gatgcggacg gcgtgcgcac catccgcctg gaaggccgcg gcaagctcac 12900
cggccaagtc atcgacgtgc cgggcgaccc gtcctcgacg gccttcccgc tggttgcggc 12960
cctgcttgtt ccgggctccg acgtcaccat cctcaacgtg ctgatgaacc ccacccgcac 13020
cggcctcatc ctgacgctgc aggaaatggg cgccgacatc gaagtcatca acccgcgcct 13080
tgccggcggc gaagacgtgg cggacctgcg cgttcgctcc tccacgctga agggcgtcac 13140
ggtgccggaa gaccgcgcgc cttcgatgat cgacgaatat ccgattctcg ctgtcgccgc 13200
cgccttcgcg gaaggggcga ccgtgatgaa cggtctggaa gaactccgcg tcaaggaaag 13260
cgaccgcctc tcggccgtcg ccaatggcct caagctcaat ggcgtggatt gcgatgaggg 13320
cgagacgtcg ctcgtcgtgc gtggccgccc tgacggcaag gggctcggca acgcctcggg 13380
cgccgccgtc gccacccatc tcgatcaccg catcgccatg agcttcctcg tcatgggcct 13440
cgtgtcggaa aaccctgtca cggtggacga tgccacgatg atcgccacga gcttcccgga 13500
gttcatggac ctgatggccg ggctgggcgc gaagatcgaa ctctccgata cgaaggctgc 13560
ctgatgagct cgaattcccg atcgttcaaa catttggcaa taaagtttct taagattgaa 13620
tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt 13680
aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc 13740
gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt 13800
atcgcgcgcg gtgtcatcta tgttactaga tcggggataa cgatcaagct ataacttcgt 13860
ataatgtatg ctatacgaag ttatcgcg 13888
<210>220
<211>866
<212>DNA
<213>大麦(Hordeum vulgare)
<400>220
ggtttacata tttaaaggaa aaaatgttca tcgattttaa aaaaaagttc atacctttca 60
aaaacgttca tcaatttttt taaaaagttc actaaaaatg aaaaaaagtt catccaattt 120
aaaaaaagtt cattaatttt ttataaattt caataaattt gaaaaaactt cataaaattc 180
aacaaaagtt cattgaagtg aaaaaagctc ataaatcttt aaaaagtgca tccattttca 240
aaaagatgtt catcaaaatt caatatagtt caccaatatt caaaaaagtt cattaatctt 300
aaaaaatatt cgctaaaatt taaaaaatgt ttatcaatat ttaaacggcg tctagatgag 360
ccggtctatt tacaaacacc ataggcgcca attaacaaaa atgcacgtta gatcacgtct 420
acggcgtcaa ataggaaatg cccatcggcc ttactattaa gagttgtttt ggttatcctt 480
taggatttat gctgtgggct ggacttaaca caaaacccac agccatggta ggccggaatc 540
tattattcag ctcacaaacg atgttctact caaaagaaga aaaaaatctg ttgtcagaaa 600
aagagaacaa aaaaggctca cgaacatgcc gcggctcgca caggtggccg tgagcttctg 660
aatgacttgg ccacccggca tgtccactgc ccccctagac ggtgtgggtg ggtggacagg 720
tcaagcgcat tgaacaaggt caccctgcgt tctgccacga ggccaactgc gtggccctca 780
tgcaacgcgc cttgctgcca cttctacaca cgccctcgcc ggccgaccgc tgctataaaa 840
gcagctcccc gttgcgtcct cgacgg 866
<210>221
<211>392
<212>DNA
<213>玉蜀黍(Zea mays)
<400>221
gatctcgtca atttttccct tttgtaattt tgtaatttat aatttacttt cctgtatgct 60
tctaactctg tcgttttaaa attttaataa aatttcagta ggggcttgcc cctcctgtcc 120
tataaaaaaa agttgacgtg aatagatttc attaagaggt tggatgttag tgggatgaca 180
tgactattag taggacgaga tgatgtggaa agttagtggg agatgatatg gatagttttt 240
gctttcatcg aaaggttgga agttagtatg atgacatggc taatatagat acatagatat 300
agactaccaa catggctgca tgcccccaag ctctcccact atatatatct ctggtagcac 360
atcatcccaa ttcacaatgc ttacaaaaac cc 392
<210>222
<211>1041
<212>DNA
<213>玉蜀黍(Zea mays)
<400>222
gtcccgcgtc aatattatta aaaaactcct acatttcttt ataatcaacc cgcactctta 60
taatctcttc tctactacta taataagaga gtttatgtac aaaataaggt gaaattatgt 120
ataagtgttc tggatattgg ttgttggctc catattcaca caacctaatc aatagaaaac 180
atatgtttta ttaaaacaaa atttatcata tatcatatat atatatatac atatatatat 240
atatatatat aaaccgtagc aatgcacggg catataacta gtgcaactta atacatgtgt 300
gtattaagat gaataagagg gtatccaaat aaaaaacttg ttcgcttacg tctggatcga 360
aaggggttgg aaacgattaa atctcttcct agtcaaaatt gaatagaagg agatttaatc 420
tctcccaatc cccttcgatc atccaggtgc aaccgtataa gtcctaaagt ggtgaggaac 480
acgaaacaac catgcattgg catgtaaagc tccaagaatt tgttgtatcc ttaacaactc 540
acagaacatc aaccaaaatt gcacgtcaag ggtattgggt aagaaacaat caaacaaatc 600
ctctctgtgt gcaaagaaac acggtgagtc atgccgagat catactcatc tgatatacat 660
gcttacagct cacaagacat tacaaacaac tcatattgca ttacaaagat cgtttcatga 720
aaaataaaat aggccggaca ggacaaaaat ccttgacgtg taaagtaaat ttacaacaaa 780
aaaaaagcca tatgtcaagc taaatctaat tcgttttacg tagatcaaca acctgtagaa 840
ggcaacaaaa ctgagccacg cagaagtaca gaatgattcc agatgaacca tcgacgtgct 900
acgtaaagag agtgacgagt catatacatt tggcaagaaa ccatgaagct gcctacagcc 960
gtctcggtgg cataagaaca caagaaattg tgttaattaa tcaaagctat aaataacgct 1020
cgcatgcctg tgcacttctc c 1041
<210>223
<211>1401
<212>DNA
<213>玉蜀黍(Zea mays)
<400>223
aagcttgccg agtgccatcc ttggacactc gataaagtat attttatttt ttttattttg 60
ccaaccaaac tttttgtggt atgttcctac actatgtaga tctacatgta ccattttggc 120
acaattacat atttacaaaa atgttttcta taaatattag atttagttcg tttatttgaa 180
tttcttcgga aaattcacat ttaaactgca agtcactcga aacatggaaa accgtgcatg 240
caaaataaat gatatgcatg ttatctagca caagttacga ccgatttcag aagcagacca 300
gaatcttcaa gcaccatgct cactaaacat gaccgtgaac ttgttatcta gttgtttaaa 360
aattgtataa aacacaaata aagtcagaaa ttaatgaaac ttgtccacat gtcatgatat 420
catatataga ggttgtgata aaaatttgat aatgtttcgg taaagttgtg acgtactatg 480
tgtagaaacc taagtgacct acacataaaa tcatagagtt tcaatgtagt tcactcgaca 540
aagactttgt caagtgtccg ataaaaagta ctcgacaaag aagccgttgt cgatgtactg 600
ttcgtcgaga tctctttgtc gagtgtcaca ctaggcaaag tctttacgga gtgtttttca 660
ggctttgaca ctcggcaaag cgctcgattc cagtagtgac agtaatttgc atcaaaaata 720
gctgagagat ttaggccccg tttcaatctc acgggataaa gtttagcttc ctgctaaact 780
ttagctatat gaattgaagt gctaaagttt agtttcaatt accaccatta gctctcctgt 840
ttagattaca aatggctaaa agtagctaaa aaatagctgc taaagtttat ctcgcgagat 900
tgaaacaggg ccttaaaatg agtcaactaa tagaccaact aattattagc tattagtcgt 960
tagcttcttt aatctaagct aaaaccaact aatagcttat ttgttgaatt acaattagct 1020
caacggaatt ctctgttttt ctaaaaaaaa actgcccctc tcttacagca aattgtccgc 1080
tgcccgtcgt ccagatacaa tgaacgtacc tagtaggaac tcttttacac gctcggtcgc 1140
tcgccgcgga tcggagtccc cggaacacga caccactgtg gaacacgaca aagtctgctc 1200
agaggcggcc acaccctggc gtgcaccgag ccggagcccg gataagcacg gtaaggagag 1260
tacggcggga cgtggcgacc cgtgtgtctg ctgccacgca gccttcctcc acgtagccgc 1320
gcggccgcgc cacgtaccag ggcccggcgc tggtataaat gcgcgccacc tccgctttag 1380
ttctgcatac agccaaccca a 1401
<210>224
<211>1805
<212>DNA
<213>薏苡(Coix lacryma-jobi)
<400>224
ggcggaagtt ttgcggcgga gctccggtcg gtgttggaga agacgaccac agcggtggcg 60
ttacggttca ccggagtagg ctagcgaggg ctcaacgaaa gctcaggagg cggcgggatc 120
agaagcacgg ccaatttggc cgtagacggc gtaacgacgg cgaggtgtgg aagcggagct 180
tggcgatttc agaggagcag aggaggaaaa cggggatggt accggcgatg aggaggctta 240
tatagccggc caaggacctg gggcgaaggg cacgtcaaag gcggtcacca cagtgcaagc 300
aggaggtggc ctggtggcgt ggcatggtcg gcagaggata cggcggtgtg gcggccgacg 360
agcagagcac gactctgctc tgatattttt tcccctccgg ttactgttca ccagagaaca 420
tctagatctt ctttccccta tttctccaaa aacattatac aacaggaaaa actcccaaca 480
tgaaagttgt tccaaaaatt aggacctaca actttcattt aaggagcaca accaaaatct 540
gcctagattt taaaatcaca ttttgaattt gaaaaacatt caaacttgaa ttcaaaatta 600
cttttgaatt ttctgaacga cttcaaattt tatcaatata aaagttgttc ctcattaaat 660
tatctacaac tttgtttttg gtcatatatc caaattgtgc ttaattttga aactaaaaaa 720
gggataaaac taggttttgg gaattgattt tcaatttgaa accaaatcaa acttgttttc 780
caaatcactt cagattctct ttagtaactt gaaaatctat cttaataaaa gttgtttctt 840
ttaatcctct ctacaacttt gattaaaaga cccaagtcta attcttttta gtttttgaaa 900
tctagtttag gggtcaaatt cagggtttga aacatttcaa atttattcaa aattttgtac 960
agaaacttta aaaactttga ataccaaagt tgtacatctc aacaagagct acaactttgc 1020
ttttgaaatc attttgaaat ttgacatact ttttgactta ttaaaaaggg gacaaaaaga 1080
gagatttaaa aaccaaggtt ttctcacttg ccctatcatg taacaccaaa catctcaatc 1140
aagccacata tatatcaaac atggaaatca acactcaaaa acatacaaca ctcttgtcaa 1200
acatacaaag catcatattt atgccttaaa cataaaaaca tgaaatgctt ttatgccatg 1260
atgccagatg attatgcata ctaaacaact taattaaaat tttaattaat gtttaacacc 1320
aggggtgtta cattaggcaa ttagtccatg ccaaacattt ctattgcatc cagagaccag 1380
aatagctttt tttttaatat ttaaccaaaa aaagaagaaa atgaggtgaa tgaatgggcc 1440
gacgggcaca taaactattg catggaccca gactattgaa ggcccgctaa tgttgagaca 1500
cggaacgcaa aaggaaaaga gggcccagac tacggcccac gcgtgggatt cggcccggcc 1560
acctccggca accagcggct catcatccaa cgccactacg ccagggcgtt cgtccacaac 1620
ctcctcgtac atatcgccgg gcggaagcgg cgcgaccgcg caagcgcaag cttgtcctcg 1680
acagcccgca caggtgtcgc gcggccccgg acacgagtcc cgcatgcgcc ccacgcggcc 1740
gcgccaggtc ccgcctcccc gcatccccac gccctctata aaccccgcgc tctccctggc 1800
cctcg 1805
<210>225
<211>232
<212>DNA
<213>稻(Oryza sativa)
<400>225
gttggcaatg cggataaaga ataactaaat aaataaataa ataaattgca agcaattgcg 60
ttgctgctat gtactgtaaa agtttcttat aatatcagtt ctgaatgcta aggacatccc 120
tcaagatggt ctttctattt ttgtgttccc gttccaatgt actgttggta tcctcttgga 180
gattcatcaa tatgagaaaa cagagaatgg acaaccctcc cttatcttat gg 232
<210>226
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>226
atgagtataa gaatgctgtt ctgc 24
<210>227
<211>21
<212>DNA
<213>人工序列
<220>
<223>引物
<400>227
gcacactgtt tttcacttaa c 21
<210>228
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>228
tcgcgcccct caagtgtcaa tacc 24
<210>229
<211>24
<212>DNA
<213>人工序列
<220>
<223>引物
<400>229
caccgcctac atacctcgct ctgc 24
<210>230
<211>16
<212>DNA
<213>人工序列
<220>
<223>引物
<400>230
ggacagcccc tcaaat 16
<210>231
<211>20
<212>DNA
<213>人工序列
<220>
<223>引物
<400>231
cctacatacc tcgctctgct 20
Claims (13)
1.一种包含编码与单体邻氨基苯甲酸合酶融合的叶绿体转运肽的多核苷酸分子的表达载体,其中:
a)所述叶绿体转运肽由SEQ ID NO:21、22、28、32、33或36的多肽序列组成;或
b)编码所述叶绿体转运肽的多核苷酸分子由SEQ ID NO:2、3、9、13、14或17组成;
其中所述叶绿体转运肽提供当与所述邻氨基苯甲酸合酶经由CTP1的靶向相比较时,所述单体邻氨基苯甲酸合酶对于植物细胞的叶绿体增强的靶向,并且其中当与具有相同基因型但缺乏所述多核苷酸分子的植物细胞相比较时,所述多核苷酸分子提供色氨酸在包含所述多核苷酸分子的转基因植物细胞中增强的水平。
2.权利要求1的表达载体,其中所述单体邻氨基苯甲酸合酶对于色氨酸是反馈不敏感的。
3.权利要求2的表达载体,其中所述单体邻氨基苯甲酸合酶由选自SEQ ID NOs:200、202、204、206、208、210和212的多肽序列组成。
4.权利要求1的表达载体,其定义为由SEQ ID NO:213或SEQ ID NO:214的核酸序列组成。
5.权利要求2的表达载体,其进一步包含与所述多核苷酸可操作地连接的在植物中起作用的启动子,所述多核苷酸编码与单体邻氨基苯甲酸合酶融合的叶绿体转运肽。
6.权利要求5的表达载体,其中所述启动子是组成型、诱导型、种子特异性或组织优选的启动子。
7.权利要求5的表达载体,其中编码叶绿体转运肽的所述多核苷酸由SEQ ID NO:9组成,并且编码单体邻氨基苯甲酸合酶的所述多核苷酸由SEQ ID NO:207组成。
8.一种转基因细胞,其用权利要求1的表达载体转化,其中所述转基因细胞不是转基因植物细胞。
9.一种增加单子叶植物的玉蜀黍种子中的游离色氨酸水平的方法,其包括在植物细胞中表达权利要求1的表达载体,其中所述叶绿体转运肽能够使所述单体邻氨基苯甲酸合酶区室化至所述植物细胞 中的质体。
10.一种产生营养上增强的玉米饲料产物的方法,其包括将用权利要求1的表达载体转化的转基因植物的种子加工成膳食、蛋白质或油,其中所述种子包含所述表达载体。
11.一种饲料产物,其通过权利要求10的方法产生。
12.权利要求11的饲料产物,其包含编码选自SEQ ID NO:21、22、28、32、33、36的多肽序列的多核苷酸分子。
13.权利要求11的饲料产物,其定义为包含当在DNA扩增法中测试时,产生对于权利要求1的表达载体诊断性的扩增子的核酸。
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US60/837,200 | 2006-08-11 | ||
PCT/US2007/075614 WO2008021974A1 (en) | 2006-08-11 | 2007-08-09 | Production of high tryptophan maize by chloroplast targeted expression of anthranilate synthase |
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IN2014DN02042A (zh) | 2011-10-28 | 2015-05-15 | Du Pont | |
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EP3270685B1 (en) | 2015-03-19 | 2023-07-19 | Pioneer Hi-Bred International, Inc. | Methods and compositions for accelerated trait introgression |
BR112018007796A2 (pt) | 2015-11-06 | 2018-10-30 | Du Pont | plantas de soja, partes de plantas de soja ou sementes de soja, método para selecionar uma célula de soja, métodos de seleção de uma célula de soja e de produção de um locus e molécula de ácido nucleico |
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- 2007-08-09 JP JP2009524003A patent/JP5296684B2/ja active Active
- 2007-08-09 CN CN200780037918.1A patent/CN101522902B/zh active Active
- 2007-08-09 AU AU2007286176A patent/AU2007286176B2/en active Active
- 2007-08-09 KR KR1020097004951A patent/KR101447300B1/ko active IP Right Grant
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
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US6248876B1 (en) * | 1990-08-31 | 2001-06-19 | Monsanto Company | Glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthases |
US5545545A (en) * | 1993-04-27 | 1996-08-13 | Regents Of The University Of Minnesota | Lysine-insensitive maize dihydrodipicolinic acid synthase |
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AU2007286176B2 (en) | 2012-10-11 |
US20120246762A1 (en) | 2012-09-27 |
US8981188B2 (en) | 2015-03-17 |
NZ574568A (en) | 2011-04-29 |
CA2660171C (en) | 2017-05-16 |
MX2009001573A (es) | 2009-02-19 |
AU2007286176A1 (en) | 2008-02-21 |
US20080050506A1 (en) | 2008-02-28 |
KR20090058519A (ko) | 2009-06-09 |
CA2660171A1 (en) | 2008-02-21 |
JP2010500035A (ja) | 2010-01-07 |
KR101447300B1 (ko) | 2014-10-07 |
WO2008021974A1 (en) | 2008-02-21 |
BRPI0716657A2 (pt) | 2013-12-03 |
JP5296684B2 (ja) | 2013-09-25 |
US8138393B2 (en) | 2012-03-20 |
EP2049670A1 (en) | 2009-04-22 |
CN101522902A (zh) | 2009-09-02 |
BRPI0716657B1 (pt) | 2018-04-17 |
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