具体实施方式
本发明人经过广泛而深入的研究,首次提供糖基转移酶gGT29-7(SEQ ID NO.:61)及其衍生多肽,如gGT29(SEQ ID NO.:26),gGT29-3(SEQ ID NO.:28),gGT29-4(SEQ IDNO.:55),gGT29-5(SEQ ID NO.:57),gGT29-6(SEQ ID NO.:59),gGT29-8(SEQ ID NO.:72),gGT29-9(SEQ ID NO.:74),gGT29-10(SEQ ID NO.:76),gGT29-11(SEQ ID NO.:78),gGT29-12(SEQ ID NO.:80),gGT29-13(SEQ ID NO.:82)gGT29-14(SEQ ID NO.:84)、gGT29-15(SEQID NO.:86)、gGT29-16(SEQ ID NO.:88)、gGT29-17(SEQ ID NO.:90)、gGT29-18(SEQ IDNO.:92)、gGT29-7-N343G(SEQ ID NO.:93)、gGT29-7-A359P(SEQ ID NO.:94)、gGT29-7-N343G/A359P(SEQ ID NO.:95)在萜类化合物糖基化催化及新皂苷合成中的应用。具体地,本发明的糖基转移酶能特异和高效地催化四环三萜化合物底物的和/或将来自糖基供体的糖基转移到四环三萜类化合物的C-3位或C-6的第一个糖基上以延伸糖链。特别是能够将人参Rh2转化为具有抗癌活性的稀有人参皂苷Rg3,将人参皂苷F2转化为人参皂苷Rd,将人参皂苷Rh1转化为抗肿瘤,抗疲劳功效的稀有人参皂苷Rf,将人参皂苷Rh1转化具有神经保护作用和紫外线保护作用的稀有人参皂苷Rg2。本发明还提供了转化和催化方法。本发明的糖基转移酶还可与达玛烯二醇和/或原人参二醇或原人参三醇合成代谢途径中的关键酶以及四环三萜C-3或者C-6位的糖基转移酶在宿主细胞中共表达,或者应用于制备人参皂苷Rh2和人参皂苷Rh1的遗传工程细胞中,应用于构建人工合成稀有人参皂苷Rg3和Rf。此外,本发明的糖基转移酶还可与达玛烯二醇和/或原人参二醇或原人参三醇合成代谢途径中的关键酶以及C-6位的糖基转移酶以及合成UDP-鼠李糖的关键酶在宿主细胞中共表达,应用于构建人工合成稀有人参皂苷Rg2的菌株。在此基础上完成了本发明。
定义
如本文所用,术语“活性多肽”、“本发明的多肽及其衍生多肽”、“本发明的酶”、“糖基转移酶”、“本发明的gGT29或其衍生多肽”或“本发明的糖基转移酶”,均指糖基转移酶gGT29-7(SEQ ID NO.:61)或其衍生多肽。其中优选的衍生多肽包括gGT29(SEQ ID NO.:26)、gGT29-3(SEQ ID NO.:28)、gGT29-4(SEQ ID NO.:55)、gGT29-5(SEQ ID NO.:57)、gGT29-6(SEQ ID NO.:59)、gGT29-8(SEQ ID NO.:72)、gGT29-9(SEQ ID NO.:74),gGT29-10(SEQ ID NO.:76),gGT29-11(SEQ ID NO.:78),gGT29-12(SEQ ID NO.:80),gGT29-13(SEQID NO.:82)gGT29-14(SEQ ID NO.:84)、gGT29-15(SEQ ID NO.:86)、gGT29-16(SEQ IDNO.:88)、gGT29-17(SEQ ID NO.:90)、gGT29-18(SEQ ID NO.:92)、gGT29-7-N343G(SEQ IDNO.:93)、gGT29-7-A359P(SEQ ID NO.:94)、gGT29-7-N343G/A359P(SEQ ID NO.:95)。
如非特别说明,本文所说的人参皂苷和皂苷元,是的C20位S和/或R构型的人参皂苷和皂苷元。
如本文所用,“分离的多肽”是指所述多肽基本上不含天然与其相关的其它蛋白、脂类、糖类或其它物质。本领域的技术人员能用标准的蛋白质纯化技术纯化所述多肽。基本上纯的多肽在非还原聚丙烯酰胺凝胶上能产生单一的主带。所述多肽的纯度还可以用氨基酸序列进行进一步分析。
本发明的活性多肽可以是重组多肽、天然多肽、合成多肽。本发明的多肽可以是天然纯化的产物,或是化学合成的产物,或使用重组技术从原核或真核宿主(例如,细菌、酵母、植物)中产生。根据重组生产方案所用的宿主,本发明的多肽可以是糖基化的,或可以是非糖基化的。本发明的多肽还可包括或不包括起始的甲硫氨酸残基。
本发明还包括所述多肽的片段、衍生物和类似物。如本文所用,术语“片段”、“衍生物”和“类似物”是指基本上保持所述多肽相同的生物学功能或活性的多肽。
本发明的多肽片段、衍生物或类似物可以是(i)有一个或多个保守或非保守性氨基酸残基(优选保守性氨基酸残基)被取代的多肽,而这样的取代的氨基酸残基可以是也可以不是由遗传密码编码的,或(ii)在一个或多个氨基酸残基中具有取代基团的多肽,或(iii)成熟多肽与另一个化合物(比如延长多肽半衰期的化合物,例如聚乙二醇)融合所形成的多肽,或(iv)附加的氨基酸序列融合到此多肽序列而形成的多肽(如前导序列或分泌序列或用来纯化此多肽的序列或蛋白原序列,或与抗原IgG片段的形成的融合蛋白)。根据本文的教导,这些片段、衍生物和类似物属于本领域熟练技术人员公知的范围。
在本发明的活性多肽具有糖基转移酶活性,并且能够催化以下一种或多种反应:
(A)
其中,R1为糖基;R2和R3为OH或者H;R4为糖基或者H;R5为糖基,所述的多肽选自SEQ ID NO.:26、28、55、57、59、61、72、74、76、78、80、82、84、86、88、90、92、93、94或95或其衍生多肽。
R1-R4经取代后的化合物如下表所示:
底物 |
R1 |
R2 |
R3 |
R4 |
Rh2 |
糖基 |
H |
OH |
H |
F2 |
糖基 |
H |
OH |
糖基 |
即当R1为葡萄糖基;R2为H,R3为OH,R4为H,式(I)化合物为Rh2。
R1为葡萄糖基;R2为H,R3为OH,R4为葡萄糖基,式(I)化合物为F2。
当以UDP-葡糖糖作为糖基供体时,底物(I)化合物为Rh2,则产物式(II)化合物为Rg3;底物(I)化合物为F2,则产物式(II)化合物为Rd;
当以UDP-木糖作为糖基供体时,底物(I)化合物为Rh2,则产物式(II)化合物为3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-PPD;底物(I)化合物为F2,则产物式(II)化合物为3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-CK。
(B)
其中,R1和R2为H或者糖基,R3和R4为糖基。R3-R4-O为C6第一个糖基衍生的糖基,所述的多肽选自SEQ ID NO.:55,57,59,61,78,82,92,94或者95或其衍生多肽。
当R1和R2为H,R3为葡萄糖基时,式(III)化合物为Rh1。
当以UDP-葡糖糖作为糖基供体时,底物(IIII)化合物为Rh1,则产物式(IV)化合物为Rf;
当以UDP-鼠李糖作为糖基供体时,底物(IIII)化合物为Rh1,则产物式(IV)化合物为Rg2;
(C)
其中,R1为糖基;R2和R3为OH或者H;R4为糖基或者H;R5为糖基,R5-R1-O为C3第一个糖基衍生的糖基;R6为糖基,R6-R1-O为C3第一个糖基衍生的糖基,所述的多肽选自SEQID NO.:26、28、59、76、84、86或88或其衍生多肽。R1为两个葡萄糖基,R2为H,R3为OH,R4为H,式(V)化合物为Rg3。
R1为两个葡萄糖基,R2为H,R3为OH,R4为葡萄糖基,式(V)化合物为Rd当以UDP-木糖作为糖基供体时,底物(V)化合物为Rg3,则产物式(VI)化合物为3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-PPD;底物(V)化合物为Rd,则产物式(VI)化合物为3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-CK。
所述的多肽序列为SEQ ID NO.:61或其衍生多肽,其优选的衍生多肽26、28、55、57、59、72、74、76、78、80、82、84、86、88、90、92、93、94或95所示的多肽,该术语还包括具有与所示多肽具有相同功能的SEQ ID NO.:26、28、55、57、59、61、72、74、76、78、80、82、84、86、88、90、92、93、94或者95序列的变异形式。这些变异形式包括(但并不限于):一个或多个(通常为1-50个,较佳地1-30个,更佳地1-20个,最佳地1-10个)氨基酸的缺失、插入和/或取代,以及在C末端和/或N末端添加一个或数个(通常为20个以内,较佳地为10个以内,更佳地为5个以内)氨基酸。例如,在本领域中,用性能相近或相似的氨基酸进行取代时,通常不会改变蛋白质的功能。又比如,在C末端和/或N末端添加一个或数个氨基酸通常也不会改变蛋白质的功能。该术语还包括本发明蛋白的活性片段和活性衍生物。本发明还提供所述多肽的类似物。这些类似物与天然多肽的差别可以是氨基酸序列上的差异,也可以是不影响序列的修饰形式上的差异,或者兼而有之。这些多肽包括天然或诱导的遗传变异体。诱导变异体可以通过各种技术得到,如通过辐射或暴露于诱变剂而产生随机诱变,还可通过定点诱变法或其他已知分子生物学的技术。类似物还包括具有不同于天然L-氨基酸的残基(如D-氨基酸)的类似物,以及具有非天然存在的或合成的氨基酸(如β、γ-氨基酸)的类似物。应理解,本发明的多肽并不限于上述例举的代表性的多肽。
修饰(通常不改变一级结构)形式包括:体内或体外的多肽的化学衍生形式如乙酰化或羧基化。修饰还包括糖基化,如那些在多肽的合成和加工中或进一步加工步骤中进行糖基化修饰而产生的多肽。这种修饰可以通过将多肽暴露于进行糖基化的酶(如哺乳动物的糖基化酶或去糖基化酶)而完成。修饰形式还包括具有磷酸化氨基酸残基(如磷酸酪氨酸,磷酸丝氨酸,磷酸苏氨酸)的序列。还包括被修饰从而提高了其抗蛋白水解性能或优化了溶解性能的多肽。
本发明的gGT29-7多肽或其衍生多肽,例如优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P蛋白的氨基端或羧基端还可含有一个或多个多肽片段,作为蛋白标签。任何合适的标签都可以用于本发明。例如,所述的标签可以是FLAG、HA、HA1、c-Myc、Poly–His、Poly-Arg、Strep-TagII、AU1、EE、T7、4A6、ε、B、gE、以及Ty1。这些标签可用于对蛋白进行纯化。表1列出了其中的一些标签及其序列。
表1
为了使翻译的蛋白分泌表达(如分泌到细胞外),还可在所述gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P的氨基酸氨基末端添加上信号肽序列,如pelB信号肽等。信号肽在多肽从细胞内分泌出来的过程中可被切去。
本发明的多核苷酸可以是DNA形式或RNA形式。DNA形式包括cDNA、基因组DNA或人工合成的DNA。DNA可以是单链的或是双链的。DNA可以是编码链或非编码链。编码成熟多肽的编码区序列可以与SEQ ID NO.:60所示的编码区序列相同或者是简并的变异体。如本文所用,“简并的变异体”在本发明中是指编码具有SEQ ID NO.:61或其衍生多肽,优选为SEQID NO.:26、28、55、57、59、72、74、76、78、80、82、84、86、88、90、92、93、94或者95的蛋白质,但与SEQ ID NO.:60或其衍生蛋白的编码序列,优选为SEQ ID NO.:25、27、54、56、58、60、71、73、75、77、79、81、83、85、87、89、或91所示的序列有差别的核酸序列。
编码SEQ ID NO.:61所示多肽或其衍生多肽,优选为SEQ ID NO.:26、28、55、57、59、72、74、76、78、80、82、84、86、88、90、92、93,94或95的成熟多肽的多核苷酸包括:只编码成熟多肽的编码序列;成熟多肽的编码序列和各种附加编码序列;成熟多肽的编码序列(和任选的附加编码序列)以及非编码序列。
术语“编码多肽的多核苷酸”可以是包括编码此多肽的多核苷酸,也可以是还包括附加编码和/或非编码序列的多核苷酸。
本发明还涉及上述多核苷酸的变异体,其编码与本发明有相同的氨基酸序列的多肽或多肽的片段、类似物和衍生物。此多核苷酸的变异体可以是天然发生的等位变异体或非天然发生的变异体。这些核苷酸变异体包括取代变异体、缺失变异体和插入变异体。如本领域所知的,等位变异体是一个多核苷酸的替换形式,它可能是一个或多个核苷酸的取代、缺失或插入,但不会从实质上改变其编码的多肽的功能。
本发明还涉及与上述的序列杂交且两个序列之间具有至少50%,较佳地至少70%,更佳地至少80%相同性的多核苷酸。本发明特别涉及在严格条件(或严紧条件)下与本发明所述多核苷酸可杂交的多核苷酸。在本发明中,“严格条件”是指:(1)在较低离子强度和较高温度下的杂交和洗脱,如0.2×SSC,0.1%SDS,60℃;或(2)杂交时加有变性剂,如50%(v/v)甲酰胺,0.1%小牛血清/0.1%Ficoll,42℃等;或(3)仅在两条序列之间的相同性至少在90%以上,更好是95%以上时才发生杂交。并且,可杂交的多核苷酸编码的多肽与SEQ ID NO.:26、28、55、57、59、61、72、74、76、78、80、82、84、86、88、90、92、93、94或者95所示的成熟多肽有相同的生物学功能和活性。
本发明还涉及与上述的序列杂交的核酸片段。如本文所用,“核酸片段”的长度至少含15个核苷酸,较好是至少30个核苷酸,更好是至少50个核苷酸,最好是至少100个核苷酸以上。核酸片段可用于核酸的扩增技术(如PCR)以确定和/或分离编码gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P、或gGT29-7-N343G/A359P蛋白的多聚核苷酸。
本发明中的多肽和多核苷酸优选以分离的形式提供,更佳地被纯化至均质。
本发明的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P核苷酸全长序列或其片段通常可以用PCR扩增法、重组法或人工合成的方法获得。对于PCR扩增法,可根据本发明所公开的有关核苷酸序列,尤其是开放阅读框序列来设计引物,并用市售的cDNA库或按本领域技术人员已知的常规方法所制备的cDNA库作为模板,扩增而得有关序列。当序列较长时,常常需要进行两次或多次PCR扩增,然后再将各次扩增出的片段按正确次序拼接在一起。
一旦获得了有关的序列,就可以用重组法来大批量地获得有关序列。这通常是将其克隆入载体,再转入细胞,然后通过常规方法从增殖后的宿主细胞中分离得到有关序列。
此外,还可用人工合成的方法来合成有关序列,尤其是片段长度较短时。通常,通过先合成多个小片段,然后再进行连接可获得序列很长的片段。
目前,已经可以完全通过化学合成来得到编码本发明蛋白(或其片段,或其衍生物)的DNA序列。然后可将该DNA序列引入本领域中已知的各种现有的DNA分子(或如载体)和细胞中。此外,还可通过化学合成将突变引入本发明蛋白序列中。
应用PCR技术扩增DNA/RNA的方法被优选用于获得本发明的基因。特别是很难从文库中得到全长的cDNA时,可优选使用RACE法(RACE-cDNA末端快速扩增法),用于PCR的引物可根据本文所公开的本发明的序列信息适当地选择,并可用常规方法合成。可用常规方法如通过凝胶电泳分离和纯化扩增的DNA/RNA片段。
本发明也涉及包含本发明的多核苷酸的载体,以及用本发明的载体或gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P蛋白编码序列经基因工程产生的宿主细胞,以及经重组技术产生本发明所述多肽的方法。
通过常规的重组DNA技术,可利用本发明的多聚核苷酸序列可用来表达或生产重组的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P多肽。一般来说有以下步骤:
(1).用本发明的编码gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P多肽的多核苷酸(或变异体),或用含有该多核苷酸的重组表达载体转化或转导合适的宿主细胞;
(2).在合适的培养基中培养的宿主细胞;
(3).从培养基或细胞中分离、纯化蛋白质。
本发明中,gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P多核苷酸序列可插入到重组表达载体中。术语“重组表达载体”指本领域熟知的细菌质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒或其他载体。只要能在宿主体内复制和稳定,任何质粒和载体都可以用。表达载体的一个重要特征是通常含有复制起点、启动子、标记基因和翻译控制元件。
本领域的技术人员熟知的方法能用于构建含gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P编码DNA序列和合适的转录/翻译控制信号的表达载体。这些方法包括体外重组DNA技术、DNA合成技术、体内重组技术等。所述的DNA序列可有效连接到表达载体中的适当启动子上,以指导mRNA合成。这些启动子的代表性例子有:大肠杆菌的lac或trp启动子;λ噬菌体PL启动子;真核启动子包括CMV立即早期启动子、HSV胸苷激酶启动子、早期和晚期SV40启动子、反转录病毒的LTRs和其他一些已知的可控制基因在原核或真核细胞或其病毒中表达的启动子。表达载体还包括翻译起始用的核糖体结合位点和转录终止子。
此外,表达载体优选地包含一个或多个选择性标记基因,以提供用于选择转化的宿主细胞的表型性状,如真核细胞培养用的二氢叶酸还原酶、新霉素抗性以及绿色荧光蛋白(GFP),或用于大肠杆菌的四环素或氨苄青霉素抗性。
包含上述的适当DNA序列以及适当启动子或者控制序列的载体,可以用于转化适当的宿主细胞,以使其能够表达蛋白质。
宿主细胞可以是原核细胞,如细菌细胞;或是低等真核细胞,如酵母细胞;或是高等真核细胞,如哺乳动物细胞。代表性例子有:大肠杆菌,链霉菌属;鼠伤寒沙门氏菌的细菌细胞;真菌细胞如酵母;植物细胞;果蝇S2或Sf9的昆虫细胞;CHO、COS、293细胞、或Bowes黑素瘤细胞的动物细胞等。
本发明的多核苷酸在高等真核细胞中表达时,如果在载体中插入增强子序列时将会使转录得到增强。增强子是DNA的顺式作用因子,通常大约有10到300个碱基对,作用于启动子以增强基因的转录。可举的例子包括在复制起始点晚期一侧的100到270个碱基对的SV40增强子、在复制起始点晚期一侧的多瘤增强子以及腺病毒增强子等。
本领域一般技术人员都清楚如何选择适当的载体、启动子、增强子和宿主细胞。
用重组DNA转化宿主细胞可用本领域技术人员熟知的常规技术进行。当宿主为原核生物如大肠杆菌时,能吸收DNA的感受态细胞可在指数生长期后收获,用CaCl2法处理,所用的步骤在本领域众所周知。另一种方法是使用MgCl2。如果需要,转化也可用电穿孔的方法进行。当宿主是真核生物,可选用如下的DNA转染方法:磷酸钙共沉淀法,常规机械方法如显微注射、电穿孔、脂质体包装等。
获得的转化子可以用常规方法培养,表达本发明的基因所编码的多肽。根据所用的宿主细胞,培养中所用的培养基可选自各种常规培养基。在适于宿主细胞生长的条件下进行培养。当宿主细胞生长到适当的细胞密度后,用合适的方法(如温度转换或化学诱导)诱导选择的启动子,将细胞再培养一段时间。
在上面的方法中的重组多肽可在细胞内、或在细胞膜上表达、或分泌到细胞外。如果需要,可利用其物理的、化学的和其它特性通过各种分离方法分离和纯化重组的蛋白。这些方法是本领域技术人员所熟知的。这些方法的例子包括但并不限于:常规的复性处理、用蛋白沉淀剂处理(盐析方法)、离心、渗透破菌、超处理、超离心、分子筛层析(凝胶过滤)、吸附层析、离子交换层析、高效液相层析(HPLC)和其它各种液相层析技术及这些方法的结合。
应用
本发明涉及的活性多肽或肽基转移酶gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P的用途包括(但不限于):特异和高效地将来自糖基供体的糖基转移到四环三萜类化合物的C-3位和C-6位的第一个糖基上以延伸糖链。特别是能够将人参皂苷Rh2转化为抗癌活性更优良的稀有人参皂苷Rg3;将人参皂苷F2转化为人参皂苷Rd;将人参皂苷Rh1转化为具有抗肿瘤和抗疲劳活性的稀有人参皂苷Rf;将人参皂苷Rh1转化具有神经保护作用和紫外线保护作用的稀有人参皂苷Rg2;本发明的糖基转移酶还能将人参皂苷Rh2,Rg3和Rd合成之前未见报道的新皂苷3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-PPD和(3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-CK。
所述的四环三萜化合物包括(但不限于):S构型或R构型的达玛烷型、羊毛脂烷型、甘遂烷型、环阿屯烷(环阿尔廷烷)型、apotirucallane型、葫芦烷、楝烷型等四环三萜类化合物。
本发明提供了一种工业催化方法,包括:在提供糖基供体的条件下,用本发明的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P活性多肽或肽基转移酶获得式(II)、(IV)或式(VI)化合物。具体是,所述的(A)反应中所用的多肽选自SEQ IDNO.:61所示多肽或其衍生多肽,优选为SEQ ID NO.:26、28、55、57、59、72、74、76、78、80、82、84、86、88、90、92、93、94或95所示氨基酸序列的活性多肽;所述(B)反应中所用的多肽选自SEQ ID NO.:55,57,59,61,78,82,92,94或者95;所述(C)反应中所用的多肽选自SEQ IDNO.:61所示多肽或其衍生多肽,优选为SEQ ID NO.:26、28、59、76、84、86或88。
所述的糖基供体是核苷二磷酸糖,选自下组:UDP-葡萄糖,ADP-葡萄糖,TDP-葡萄糖,CDP-葡萄糖,GDP-葡萄糖,UDP-乙酰基葡萄糖,ADP-乙酰基葡萄糖,TDP-乙酰基葡萄糖,CDP-乙酰基葡萄糖,GDP-乙酰基葡萄糖,UDP-木糖,ADP-木糖,TDP-木糖,CDP-木糖,UDP-木糖,GDP-木糖,UDP-半乳糖醛酸,ADP-半乳糖醛酸,TDP-半乳糖醛酸,CDP-半乳糖醛酸,GDP-半乳糖醛酸,UDP-半乳糖,ADP-半乳糖,TDP-半乳糖,CDP-半乳糖,GDP-半乳糖,UDP-阿拉伯糖,ADP-阿拉伯糖,TDP-阿拉伯糖,CDP-阿拉伯糖,GDP-阿拉伯糖,UDP-鼠李糖,ADP-鼠李糖,TDP-鼠李糖,CDP-鼠李糖,GDP-鼠李糖,或其他核苷二磷酸己糖或核苷二磷酸戊糖,或其组合。
所述的糖基供体优选是尿苷二磷酸糖,选自下组:UDP-葡萄糖,UDP-木糖,UDP-鼠李糖,UDP-半乳糖醛酸,UDP-半乳糖,UDP-阿拉伯糖,或其他尿苷二磷酸己糖或尿苷二磷酸戊糖,或其组合。
在所述方法中,还可以添加酶活性添加物(提高酶活性或抑制酶活性的添加物)。所述酶活性的添加物可以选自下组:Ca2+、Co2+、Mn2+、Ba2+、Al3+、Ni2+、Zn2+、或Fe2+;或为可以生成Ca2+、Co2+、Mn2+、Ba2+、Al3+、Ni2+、Zn2+、或Fe2+的物质。
所述方法的pH条件为:pH4.0-10.0,优选pH6.0-pH8.5,更优选8.5。
所述方法的温度条件为:10℃-105℃,优选25℃-35℃,更优选35℃。
本发明还提供了一种组合物,它含有有效量的本发明的活性多肽或肽基转移酶gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或者gGT29-7-N343G/A359P,以及食品学上或工业上可接受的载体或赋形剂。这类载体包括(但并不限于):水、缓冲液、葡萄糖、水、甘油、乙醇、及其组合。
所述的组合物中还可添加调节本发明糖基转移酶活性的物质。任何具有提高酶活性功能的物质均是可用的。较佳地,所述的提高本发明的糖基转移酶活性的物质选自巯基乙醇。此外,很多物质可以降低酶活性,包括但不限于:Ca2+、Co2+、Mn2+、Ba2+、Al3+、Ni2+、Zn2+和Fe2+;或在添加至底物后可水解形成Ca2+、Co2+、Mn2+、Ba2+、Al3+、Ni2+、Zn2+和Fe2+的物质。
在获得了本发明的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P后,本领域人员可以方便地应用该酶来发挥转糖基的作用,特别是对达玛稀二醇、原人参二醇和原人参三醇的转糖基作用。作为本发明的优选方式,还提供了二种形成稀有人参皂苷的方法,该方法之一包含:用本发明所述的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P处理待转糖基的底物,所述的底物包括达玛稀二醇、原人参二醇和原人参三醇及其衍生物等四环三萜类化合物。较佳地,在pH3.5-10条件下,用所述的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G或gGT29-7-A359P或gGT29-7-N343G/A359P酶处理待转糖基的底物。较佳地,在温度30-105℃条件下,用所述的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P、gGT29-7-N343G/A359P、gGT29-3酶处理待转糖基的底物。
该方法之二包含:将本发明所述的gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P或gGT29-7-N343G/A359P基因转入可以合成人参皂苷Rh2或者Rh1的工程菌(例如,酵母或大肠杆菌工程菌)中,或者,将gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、gGT29-7-A359P、或gGT29-7-N343G/A359P基因与达玛稀二醇、原人参二醇和原人参三醇合成代谢途径中的关键基因以及四环三萜C-3和/或C-6位的糖基转移酶于宿主细胞(例如酵母细胞或大肠杆菌)中共表达,获得直接生产稀有人参皂苷Rg3或Rf的重组菌。或者,将gGT29-7多肽或其衍生多肽,优选为gGT29、gGT29-3、gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18、gGT29-7-N343G、或gGT29-7-A359P、或gGT29-7-N343G/A359P基因与达玛烯二醇和/或原人参二醇或原人参三醇合成代谢途径中的关键酶以及四环三萜C-6位的糖基转移酶以及合成UDP-鼠李糖的关键酶在宿主细胞中共表达,应用于构建人工合成稀有人参皂苷Rg2的重组菌株。
所述的达玛稀二醇合成代谢途径中的关键基因包括(但不限于):达玛烯二醇合成酶基因。
在另一优选例中,所述的原人参二醇合成代谢途径中的关键基因包括(但不限于):达玛烯二醇合成酶基因、细胞色素P450 CYP716A47基因及其还原酶基因,或其组合。或者以上各种酶的同功酶及其组合。其中,达玛烯二醇合成酶将环氧角鲨烯(酿酒酵母自身合成)转化为达玛烯二醇,细胞色素P450 CYP716A47及其还原酶再将达玛烯二醇转化为原人参二醇。(Han et.al,plant&cell physiology,2011,52.2062-73)
在另一优选例中,所述的原人参三醇合成代谢途径中的关键基因包括(但不限于):达玛烯二醇合成酶基因、细胞色素P450 CYP716A47基因及其还原酶基因、及细胞色素P450 CYP716A53V2基因及其还原酶基因,或其组合。或者以上各种酶的同功酶及其组合。其中,达玛烯二醇合成酶将环氧角鲨烯(酿酒酵母自身合成)转化为达玛烯二醇,细胞色素P450 CYP716A47及其还原酶再将达玛烯二醇转化为原人参二醇,细胞色素P450CYP716A53v2(JII036031)及其还原酶再进一步将原人参二醇转化为原人参三醇。(Hanet.al,plant&cell physiology,2012,53.1535-45)
在另一优选例中,所述的人参皂苷Rh2合成代谢途径中的关键基因包括(但不限于):达玛烯二醇合成酶基因、细胞色素P450 CYP716A47基因及其还原酶基因、四环三萜C-3位糖基转移酶UGTPg45或其组合。或者以上各种酶的同功酶及其组合。其中,达玛烯二醇合成酶将环氧角鲨烯(酿酒酵母自身合成)转化为达玛烯二醇,细胞色素P450 CYP716A47及其还原酶再将达玛烯二醇转化为原人参二醇,糖基转移酶UGTPg45可以进一步把原人参二醇转化为Rh2(Wang et.al,Metabolic Engineering,2015,29.97-105)。
在另一优选例中,所述的人参皂苷Rh1合成代谢途径中的关键基因包括(但不限于):达玛烯二醇合成酶基因、细胞色素P450 CYP716A47基因及其还原酶基因、及细胞色素P450 CYP716A53V2基因及其还原酶基因,及C-6糖基转移酶UGTPg100或其组合。或者以上各种酶的同功酶及其组合。其中,达玛烯二醇合成酶将环氧角鲨烯(酿酒酵母自身合成)转化为达玛烯二醇,细胞色素P450CYP716A47及其还原酶再将达玛烯二醇转化为原人参二醇,细胞色素P450CYP716A53v2(JII036031)及其还原酶再进一步将原人参二醇转化为原人参三醇,糖基转移酶UGTPg100可以进一步把原人参三醇转化为Rh1(Wei et.al,MolecularPlant,2015,15.doi:10.1016/j.molp.2015.05.010)。
本发明的主要优点:
(1)本发明的糖基转移酶可以特异性和高效地将来自糖基供体的糖基转移到四环三萜类化合物的C-3位和/或C-6位的第一个糖基上以延伸糖链
(2)本发明的糖基转移酶特别能够分别将人参皂苷Rh2和Rh1转化为具有更好抗癌活性的稀有人参皂苷Rg3和具有抗肿瘤,抗疲劳的功效的稀有人参皂苷Rf以及具有神经保护中的作用和紫外线保护作用的稀有人参皂苷Rg2
(3)在酵母中构建了人参皂苷元(达玛稀二醇、原人参二醇和原人参三醇)的合成途径或稀有人参皂苷Rh2或Rh1的合成途径,从而实现以葡萄糖等单糖为底物,用酵母来发酵生产稀有人参皂苷Rg3和Rf。在酵母中构建了稀有人参皂苷Rh1的合成途径以及合成UDP-鼠李糖的途径,用酵母来发酵生产稀有人参皂苷Rg2。这不但可以解决皂苷生产的原料来源问题,而且可以大幅降低稀有皂苷Rg3、Rg2和Rf的生产成本。
下面结合具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法,通常按照常规条件如Sambrook等人,分子克隆:实验室手册(New York:Cold Spring Harbor LaboratoryPress,1989)中所述的条件,或按照制造厂商所建议的条件。
实施例1糖基转移酶及其编码基因的分离
从发表的人参属植物表达谱数据提取了100多条预测为糖基转移酶的cDNA序列,从中克隆了60条cDNA全长序列并对它们进行了表达及转糖基反应分析,其中有17种表达产物对人参皂苷元及皂苷具有转糖基活性。
提取人参RNA并进行反转录,获得人参的cDNA。以该cDNA为模板进行PCR扩增,使用其中引物对1(SEQ ID NO.:7、8);引物对2(SEQ ID NO.:9、10);引物对3(SEQ ID NO.:11,12);引物对5(SEQ ID NO.:34、35);引物对7(SEQ ID NO.:46、47);引物对8(SEQ ID NO.:62、63);引物对9(SEQ ID NO.:64、65)全部获得扩增产物。DNA聚合酶选用宝生物工程有限公司的高保真的KOD DNA聚合酶。PCR产物经琼脂糖凝胶电泳检测(图1)。在紫外下照射,切下目标DNA条带。然后采用AIIygen Gel EIItraction Kit(AEYGEN公司)从琼脂糖凝胶中回收DNA即为扩增出的DNA片段。将此DNA片段用宝生物工程有限公司的rTaq DNA聚合酶在末端加A后与市售的克隆载体pMD18-T Vector连接,连接产物转化市售的大肠杆菌EPI300感受态细胞,将转化后的大肠杆菌菌液涂布在添加氨苄青霉素50ug/mL、IPTG 0.5mM、II-Gal25μg/mL的LB平板上,并进一步通过PCR和酶切验证重组克隆。分别选取其中一个克隆提取重组质粒后进行测序。用BESTORF软件寻找开放阅读框(ORF)。通过序列比对,ORF编码了糖基转移酶第1家族保守功能域PSPG盒,表明是糖基转移酶基因。
用引物对5(SEQ ID NO.:34,35)获得基因具有SEQ ID NO.:25,27所示的核苷酸序列,分别命名为gGT29和gGT29-3。其中,相应的基因信息见表2。
表2
SEQ ID NO.: |
自5'的起始位点 |
基因名称 |
作用 |
序列 |
25 |
1-1329 |
gGT29 |
开放阅读框 |
|
25 |
1-3 |
gGT29 |
起始密码子 |
ATG |
25 |
1327-1329 |
gGT29 |
终止密码子 |
TAG |
27 |
1-1329 |
gGT29-3 |
开放阅读框 |
|
27 |
1-3 |
gGT29-3 |
起始密码子 |
ATG |
27 |
1327-1329 |
gGT29-3 |
终止密码子 |
TAG |
用引物对7(SEQ ID NO.:62、63)获得基因具有表3所示的gGT29-7衍生多肽的核苷酸序列,命名为gGT29-4、gGT29-5、gGT29-6、gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-17和gGT29-18。其中相应的基因信息见表3。
表3
用引物对8(SEQ ID NO.:64、65)获得基因具有表4所示的核苷酸序列,命名为gGT29-7、gGT29-15和gGT29-16。其中,相应的基因信息见表4。
表4
SEQ ID NO.: |
自5'的起始位点 |
基因名称 |
作用 |
序列 |
60 |
1-1341 |
gGT29-7 |
开放阅读框 |
|
60 |
1-3 |
gGT29-7 |
起始密码子 |
ATG |
60 |
1339-1341 |
gGT29-7 |
终止密码子 |
TAG |
86 |
1-1329 |
gGT29-15 |
开放阅读框 |
|
86 |
1-3 |
gGT29-15 |
起始密码子 |
ATG |
86 |
1327-1329 |
gGT29-15 |
终止密码子 |
TAG |
88 |
1-1329 |
gGT29-16 |
开放阅读框 |
|
88 |
1-3 |
gGT29-16 |
起始密码子 |
ATG |
88 |
1327-1329 |
gGT29-16 |
终止密码子 |
TAG |
所涉及的蛋白及其特性如5表所示:
表5
实施例2糖基转移酶基因gGT29和gGT29-3的酵母重组表达载体的构建
分别以实施例1构建的含有gGT29和gGT29-3基因的质粒gGT29-pMD18T和gGT29-3-pMD18T为模板扩增目标基因。
gGT29所用正向引物均为(SEQ ID NO.:36),其5’端添加Kpn I识别位点:GGATCC;所用反向引物均为(SEQ ID NO.:37),其5’端添加IIhoI识别位点:CTCGAG,反向引物引入6-His Tag以便进行Western Blot检测表达和纯化。
gGT29-3所用正向引物均为(SEQ ID NO.:38),其5’端添加Kpn I识别位点:GGATCC;所用反向引物均为(SEQ ID NO.:39),其5’端添加IIhoI识别位点:CTCGAG,反向引物引入6-His Tag以便进行Western Blot检测表达和纯化。
以质粒gGT29-pMD18T和gGT29-3-pMD18T为模板,利用上述引物通过PCR方法扩增gGT29和gGT29-3的基因。DNA聚合酶选用Toyobo公司的高保真DNA聚合酶kod,参考其说明书设定PCR程序:94℃2min;94℃15s,58℃30s,68℃1.5min,共30个循环;68℃10min;10℃保温。PCR产物经琼脂糖凝胶电泳检测,在紫外光下,切下与目标DNA大小一致的条带。然后采用AIIYGEN公司的AIIyPrep DNA Gel EIItraction Kit从琼脂糖凝胶中回收DNA片段。用Takara公司的的QuickCut限制性内切酶Kpn I和IIba I双酶切回收的DNA片段30min,用AIIYGEN公司的AIIyPrep PCR Cleanup Kit对酶切产物进行清洁回收。利用NEB公司的T4DNA连接酶将酶切产物与酿酒酵母表达质粒pYES2(同样以Kpn I和IIba I酶切并割胶回收)25℃连接2h。连接产物转化E.coli TOP 10感受态细胞,并涂布于添加100μg/mL氨苄青霉素的LB平板上。通过菌落PCR验证阳性转化子,并测序进一步验证表达质粒gGT29-pYES2和gGT29-3-pYES2构建成功。
实施例3糖基转移酶基因gGT29和gGT29-3在酿酒酵母中的表达
通过电转化方法将构建好的表达质粒gGT29-pYES2和gGT29-3-pYES2转化到酿酒酵母(Saccharomyces cerevisiae)中,涂布于筛选平板SC-Ura(0.67%酵母无氨基酸基本氮源,2%葡萄糖)。通过菌落PCR验证酵母重组子。挑酵母重组子菌落于10mL SC-Ura(2%葡萄糖)培养基中,30℃200rpm培养20h。4℃3500g离心收集菌体,用无菌去离子水清洗菌体两次,用诱导培养基SC-Ura(2%半乳糖)重悬菌体,并接种到50mL诱导培养基中,使OD600在0.4左右,30℃200rpm开始诱导表达。4℃3500g离心收集诱导表达12h的菌体,用无菌去离子水清洗菌体两次,重悬于酵母裂解缓冲液中,使OD600在50-100之间。用Fastprep细胞破碎仪震荡破碎酵母细胞,4℃12000g离心10min去除细胞碎片,收集细胞裂解液上清。取适量裂解液上清进行SDS-PAGE电泳检测,与pYES2空载体的重组子相比,gGT29-pYES2和gGT29-3-pYES2重组子没有明显的条带表征(图2)。采用anti-6-His Tag Western Blot检测表达情况,表达gGT29和gGT29-3的酿酒酵母显示很强的Western Blot信号,表明gGT29和gGT29-3在酵母中均可溶表达,而转pYES2空载体的重组子则没有anti-6-His Tag Western Blot信号(图3)。
实施例4酵母表达产物gGT29和gGT29-3转糖基反应和产物鉴定
以表达gGT29和gGT29-3的重组酵母裂解上清做为酶液来催化人参皂苷Rh2和F2的转糖基反应,表达空载体的重组酵母裂解上清为对照。100μL反应体系如表3所示。反应在35℃下进行12h,然后加入100μL丁醇终止反应,并抽提产物。产物真空干燥后,用甲醇溶解。
反应产物先用薄层层析(TLC)进行检测,表达gGT29和gGT29-3的酵母宿主裂解上清酶液可以在人参皂苷Rh2和F2的3位糖基再延伸一个糖基,转化为人参皂苷Rg3和Rd(图4)。gGT29和gGT29-3的催化活性不受人参皂苷20位糖基或者羟基构型的影响,可以将20(R)-Rh2转化为20(R)-Rg3(图6)。
实施例5糖基转移酶BvUGT73C10/UGTPg45和gGT29的联合转糖基反应和产物鉴定
以表达BvUGT73C10(JQ291613)或UGTPg45(KM401918)的大肠杆菌宿主裂解上清和以表达gGT29的酵母宿主裂解上清做为酶液来共同催化原人参二醇(PPD)。100μL反应体系如表3所示。73.4μL的酶液中40μL为BvUGT73C10的大肠宿主裂解上清,剩余33.4μL为表达gGT29的酵母宿主裂解上清。反应在35℃下进行12h,然后加入100μL丁醇终止反应,并抽提产物。产物真空干燥后,用甲醇溶解。反应产物先用薄层层析(TLC)进行检测(图5),可以看到糖基转移酶BvUGT73C10和gGT29或者UGTPg45和gGT29联合使用都可将PPD转化为Rg3。
糖基转移酶BvUGT73C10和gGT29或者3GT2和gGT29联合使用都可以催化20(R)-PPD,生成20(R)-Rg3(图6)。
实施例6产Rg3酵母工程菌的构建与产物鉴定
6.1在pESC-HIS质粒((Stratagene,Agilent)上,同时组装达玛烯二醇合成酶(Dammarenediol synthase)(ACZ71036.1)(GAL1/GAL10 GAL10侧启动子,ADH1终止子)、细胞色素P450 CYP716A47(AEY75213.1)(FBA1启动子,CYC1终止子)以及糖基转移酶UGTPg45和gGT29(GAL1/GAL10 GAL1侧启动子,TDH2终止子),构成游离型质粒,转化酿酒酵母BY4742,并将拟南芥来源的细胞色素P450还原酶ATR2-1(NP_849472.2)整合于酿酒酵母BY4742染色体中染色体trp1基因位点(GAL1启动子,利用trp1原有终止子),构建了重组酵母A2。重组酵母菌需补加的相应氨基酸(0.01%色氨酸,0.01%亮氨酸,0.01%赖氨酸)。
将重组酵母A2裂解液转移到2mL EP管中,每个管装1mL,加入等体积(1mL)的正丁醇抽提约30min后12000g离心10min。吸取上清至一新的EP管中。45℃并真空条件下使正丁醇蒸干。用100μL甲醇溶解后用于HPLC检测。
通过HPLC和LC-MS分析,重组酵母A2的细胞裂解液中含有达玛烯二醇、原人参二醇(PPD)和人参皂苷活性代谢物Rg3(图8和图9)。
6.2方法同6.1,区别在于用糖基转移酶BvUGT73C10代替UGTPg45,得到重组酵母A6。通过HPLC分析,重组酵母A6的细胞裂解液中也含有达玛烯二醇、原人参二醇(PPD)及人参皂苷活性代谢物Rg3。
实施例7糖基转移酶基因gGT29-4,gGT29-5,gGT29-6和gGT29-7的大肠杆菌重组表达载体的构建
以实施例1构建的含有gGT29-4,gGT29-5,gGT29-6和gGT29-7基因的质粒gGT29-4-pMD18T,gGT29-5-pMD18T,gGT29-6-pMD18T和gGT29-7-pMD18T为模板扩增目标基因。
gGT29-5和gGT29-6基因所用的正向引物如SEQ ID NO.:66所示,其5’端添加了与载体pET28a同源的序列:CTGGTGCCGCGCGGCAGC;所用反向引物为如SEQ IDNO.:68所示,其5’端添加了与载体pET28a同源的序列:TGCGGCCGCAAGCTTGTC。
gGT29-4和gGT29-7基因所用正向引物为SEQ ID NO.:67,其5’端添加了与载体pET28a同源的序列:CTGGTGCCGCGCGGCAGC;所用反向引物为SEQ ID NO.:68,其5’端添加了与载体pET28a同源的18个碱基片段:TGCGGCCGCAAGCTTGTC。
利用上述引物通过PCR方法扩增gGT29-4,gGT29-5,gGT29-6和gGT29-7基因。扩增基因选用NEB公司的Q5高保真DNA聚合酶,参考其说明书设定PCR程序:98℃30s;98℃15s,58℃30s,72℃1min,共35个循环;72℃2min;10℃保温。
同时,使用SEQ ID NO.:69和SEQ ID NO.:70分别作为正向和反向引物扩增载体pET28a,获得线性化的载体pET28a。扩增pET28a线性化载体也选用NEB公司的Q5高保真DNA聚合酶,参考其说明书设定PCR程序:98℃30s;98℃15s,58℃30s,72℃3min,共35个循环;72℃2min;10℃保温。
上述gGT29-4,gGT29-5,gGT29-6和gGT29-7基因PCR产物以及线性化的载体pET28a,经琼脂糖凝胶电泳检测后,在紫外光下切下与目标DNA大小一致的条带。然后采用AXYGEN公司的AxyPrep DNA Gel Extraction Kit从琼脂糖凝胶中回收DNA片段。参考诺晶生物科技有限公司的BGclonart无缝克隆试剂盒说明书,将回收的线性化的pET28a载体片段,回收的gGT29-4,gGT29-5,gGT29-6和gGT29-7基因片段及诺晶生物科技有限公司的BGclonart无缝克隆反应液以适当比例进行混合,共20μl。混匀后在50℃孵育30分种,然后将混合反应液转移到冰上。使用5μl反应液转化E.coli EPI300感受态细胞,并涂布于添加50μg/mL卡那霉素的LB平板上。通过菌落PCR验证阳性转化子,并测序进一步验证表达质粒gGT29-4-pET28a,gGT29-5-pET28a,gGT29-6-pET28a和gGT29-7-pET28a构建成功。
实施例8糖基转移酶基因gGT29-4,gGT29-5,gGT29-6和gGT29-7在大肠杆菌中的表达
以实施例19构建的大肠杆菌表达载体gGT29-4-pET28a,gGT29-5-pET28a,gGT29-6-pET28a和gGT29-7-pET28a转化到市售的E.coli BL21中。接种一个重组子到LB培养基中,30℃200rpm培养至OD600约0.6-0.8,使菌液降温至4℃,加入终浓度为50μM的IPTG,18℃200rpm诱导表达15h。4℃离心收集菌体,超声破碎细胞,4℃12000g离心收集细胞裂解液上清,取样品进行SDS-PAGE电泳(图10)。gGT29-4-pET28a,gGT29-5-pET28a,gGT29-6-pET28a和gGT29-7-pET28a重组子裂解液和总蛋白和上清中都有明显的目的蛋白条带(大约50KD),分别表征糖基转移酶gGT29-4,gGT29-5,gGT29-6和gGT29-7。从Western Blot的结果看(图11),也证明目标蛋白gGT29-4,gGT29-5,gGT29-6和gGT29-7在宿主中实现了可溶表达。
实施例9大肠杆菌表达产物gGT29-4,gGT29-5,gGT29-6和gGT29-7转糖基反应和产物的鉴定
以表达gGT29-4,gGT29-5,gGT29-6和gGT29-7的重组酵母裂解上清做为酶液来催化人参皂苷Rh2和F2的转糖基反应。100μL反应体系如表6所示。反应在35℃下进行12h,然后加入100μL丁醇终止反应,并抽提产物。产物真空干燥后,用甲醇溶解。
表6
9%吐温20 |
11.1μL |
50mM UDP-葡萄糖 |
10μL |
1M Tris-HCl pH8.5 |
5μL |
100mM底物(乙醇溶解) |
0.5μL |
酶液 |
73.4μL |
反应产物用薄层层析(TLC)进行检测,gGT29-6的粗酶液可以在人参皂苷Rh2和F2的3位糖基上再延伸一个糖基,分别生成为人参皂苷Rg3和Rd(图12);gGT29-4,gGT29-5和gGT29-7的粗酶液可以在人参皂苷F2的3位糖基再延伸一个糖基生成皂苷Rd,但是它们不能催化皂苷Rh2(图12)。gGT29-4,gGT29-5,gGT29-6和gGT29-7的粗酶液还可以在原人参三醇型皂苷Rh1的C-6位糖基上再延伸一个糖基,形成人参皂苷Rf(图13),其中,gGT29-4,gGT29-5和gGT29-6的活性比较弱,而gGT29-7则具有比较强的活性(表7)。
实施例10糖基转移酶基因gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18大肠杆菌重组表达载体的构建、在大肠杆菌中的表达以及产物鉴定
方法同实施例7-8,构建了gGT29-8、gGT29-9、gGT29-10、gGT29-11、gGT29-12、gGT29-13、gGT29-14、gGT29-15、gGT29-16、gGT29-17、gGT29-18的大肠杆菌表达载体(gGT29-8-pET28a,gGT29-9-pET28a,gGT29-10-pET28a,gGT29-11-pET28a,gGT29-12-pET28a,gGT29-13-pET28a,gGT29-14-pET28a,gGT29-15-pET28a,gGT29-16-pET28a,gGT29-17-pET28a,gGT29-18-pET28a),并在大肠杆菌中实现可溶性表达。根据实施例9所示方法的鉴定,其产物与其他gGT29系列糖基转移酶对底物不同位置(C3或C6)第一个糖基的延伸活性见表7,其中,“+"表示在该位置有活性,“++"表示在该位置有比较强的活性。
表7
实施例11通过氨基酸定点突变改变gGT29-7对糖基供体专一性
糖基转移酶gGT29-7可以以UDP-葡萄糖为糖基供体,催化人参皂苷Rh1的C6-O-Glc延伸一个葡萄糖生成人参皂苷Rf。通过对对糖基转移酶gGT29-7的糖基供体结合区域(PSPG)进行了定点突变来改变它对糖基供体的专一性。对已有研究报道的能够催化其他糖基受体的鼠李糖基转移酶与gGT29-7进行氨基酸序列比对,筛选并最后选取了gGT29-7PSPG盒上两个位点进行定点突变(N343G和A359P)。
根据常规设计并合成两条引物,以质粒pET28a-gGT29-7为模板,利用Yeasen公司的Hieff MutTM site-directed mutagenesis kit进行gGT29-7的N343位点分别进行定点突变。
PCR扩增程序为:95℃30s;95℃5s,56℃10s,72℃1.5min,共30个循环;降至10℃。PCR产物用DpnI酶切,37℃水浴反应2h。再取3μL用试剂盒中的重组酶EIInase进行重组37℃30min,转化大肠杆菌TOP 10感受态。挑取单克隆并抽提质粒,测序验证突变位点,所获得的质粒命名为pET28a-gGT29-7-N343G。
将质粒pET28a-gGT29-7-N343G转化到大肠杆菌BL21(DE3),构建重组菌株pET28a-gGT29-7-N343G-BL21,诱导表达的步骤同实施例8。
根据常规设计并合成两条引物对gGT29-7的A359位点进行点突变并构建突变质粒pET28a-gGT29-7-A359P和重组菌株gGT29-7-N343G-BL21,步骤同上。诱导表达的步骤同实施例8。以质粒pET28a-gGT29-7-N343G为模板,29-7-A359P-F和29-7-A359P-R为引物对A359进行点突变构建双突变质粒pET28a-gGT29-7-N343G/A359P和重组菌株gGT29-7-N343G/A359P-BL21,步骤同上。诱导表达的步骤同实施例8,转糖基反应和产物的鉴定同实施例9,反应体系如表6所示,但是使用按照1/1的比例混合的UDP-葡萄糖/UDP-鼠李糖代替UDP-葡萄糖作为糖基供体。
结果如图14所示,单突变蛋白gGT29-7-N343G催化Rh1的C6-O-Glc延伸一个葡萄糖生成Rf的酶活力已经基本检测不到,而gGT29-7-A359P仍然保留了催化Rh1的C6-O-Glc延伸一个葡萄糖的酶活力。双突变蛋白gGT29-7-N343G/A359P不仅保留了野生型蛋白的催化Rh1的C6-O-Glc延伸一个葡萄糖生成Rf的酶活力,还获得了催化Rh1的C6-O-Glc延伸一个鼠李糖生成Rg2的酶活力。
实施例12以UDP-xylose为糖基供体,糖基转移酶大肠杆菌表达产物的转糖基反应和产物的鉴定
以表达gGT29-10和gGT29-14的重组大肠杆菌上清做为酶液来催化人参皂苷Rh2和F2的转糖基反应。100μL反应体系如表6所示,但是用UDP-木糖代替UDP-葡萄糖作为糖基供体。反应在35℃下进行12h,然后加入100μL丁醇终止反应,并抽提产物。产物真空干燥后,用甲醇溶解。
反应产物用薄层层析(TLC)进行检测,gGT29-10和gGT29-14能够在人参皂苷Rh2的C-3糖基延伸一个木糖生成一种新的三萜皂苷,(3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-PPD;gGT29-10和gGT29-14能够将人参皂苷Rg3 C-3位的第二个葡萄糖置换为木糖生成一种新的三萜皂苷(3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-PPD);gGT29-10和gGT29-14还能够将人参皂苷Rd C-3位的第二个葡萄糖置换为木糖生成一种新的三萜皂苷(3-O-β-(D-xylopyranosyl)-β-(D-glucopyranosyl)-CK)(图15)。
其它糖基转移酶gGT29,gGT29-4,gGT29-5,gGT29-6和,gGT29-7 gGT29-8,gGT29-9,gGT29-10,gGT29-11,gGT29-12,gGT29-13,gGT29-15,gGT29-16,gGT29-17和gGT29-18,以UDP-木糖为糖基供体,催化Rh2,Rg3和Rd的活性如表8所示。
表8
实施例13产Rf酵母工程菌的构建与产物鉴定
在pESC-HIS质粒((Stratagene,Agilent)上,同时组装达玛烯二醇合成酶(Dammarenediol synthase)(ACZ71036.1)(GAL1/GAL10 GAL10侧启动子,ADH1终止子)、细胞色素P450 CYP716A47(AEY75213.1)(FBA1启动子,CYC1终止子)、细胞色素P450CYP716A53V2基因(ENO2启动子,CYC1终止子)以及糖基转移酶基因UGTPg100(KP795113)(GAL1/GAL10 GAL1侧启动子,TDH2终止子)和糖基转移酶gGT29-7(TEF1启动子,FBA1终止子),构成游离型质粒,转化酿酒酵母BY4742,并将拟南芥来源的细胞色素P450还原酶ATR2-1(NP_849472.2)整合于酿酒酵母BY4742染色体中染色体trp1基因位点(GAL1启动子,利用trp1原有终止子),构建了重组酵母A5。重组酵母菌需补加的相应氨基酸(0.01%色氨酸,0.01%亮氨酸,0.01%赖氨酸)。
将重组酵母A7裂解液转移到2mL EP管中,每个管装1mL,加入等体积(1mL)的正丁醇抽提约30min后12000g离心10min。吸取上清至一新的EP管中。45℃并真空条件下使正丁醇蒸干。用100μL甲醇溶解后用于HPLC检测。
通过HPLC分析,重组酵母A7的细胞裂解液中含有原人参三醇(PPT)及人参皂苷活性代谢物Rh1和Rf。
实施例14产Rg2酵母工程菌的构建与产物鉴定
在pESC-HIS质粒((Stratagene,Agilent)上,同时组装达玛烯二醇合成酶(Dammarenediol synthase)(ACZ71036.1)(GAL1/GAL10 GAL10侧启动子,ADH1终止子)、细胞色素P450 CYP716A47(AEY75213.1)(FBA1启动子,CYC1终止子)、细胞色素P450CYP716A53V2基因(ENO2启动子,CYC1终止子)以及两个糖基转移酶基因UGTPg100(KP795113)(GAL1/GAL10 GAL1侧启动子,TDH2终止子)和gGT29-7-N343G/A359P(TEF1启动子,FBA1终止子),构成游离型质粒,转化酿酒酵母BY4742,并将拟南芥来源的细胞色素P450还原酶ATR2-1(NP_849472.2)(GAL1启动子,利用trp1原有终止子)和UDP-L-鼠李糖合成酶RHM2(GQ292791.1)(GAL10启动子,FBA1终止子)整合于酿酒酵母BY4742染色体中染色体trp1基因位点,构建了重组酵母A8。重组酵母菌需补加的相应氨基酸(0.01%色氨酸,0.01%亮氨酸,0.01%赖氨酸)。
将重组酵母A8裂解液转移到2mL EP管中,每个管装1mL,加入等体积(1mL)的正丁醇抽提约30min后12000g离心10min。吸取上清至一新的EP管中。45℃并真空条件下使正丁醇蒸干。用100μL甲醇溶解后用于HPLC检测。通过HPLC分析,重组酵母A8的细胞裂解液中含有原人参三醇(PPT)及人参皂苷活性代谢物Rh1和Rg2。
在本发明提及的所有文献都在本申请中引用作为参考,就如同每一篇文献被单独引用作为参考那样。此外应理解,在阅读了本发明的上述讲授内容之后,本领域技术人员可以对本发明作各种改动或修改,这些等价形式同样落于本申请所附权利要求书所限定的范围。
序列表
<110> 周志华
<120> 一组糖基转移酶及其应用
<130> P2015-0697
<150> CN201410254021.6
<151> 2014-06-09
<160> 95
<170> PatentIn version 3.5
<210> 1
<211> 1425
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25
<400> 1
atgaagtcag aattgatatt cttgcccgcc ccggccatcg gacacctcgt gggaatggtg 60
gagatggcta aactcttcat cagtcgacat gaaaacctct cggtcaccgt cctcatcgcg 120
aaattctaca tggatacggg ggtagacaac tacaataaat cactcttaac aaaccctacc 180
ccgcgtctca caattgtaaa tctcccggaa accgaccccc aaaactatat gctcaaacca 240
cgccatgcca tctttcctag cgtcatcgag actcagaaga cacacgtgcg agacataata 300
tcaggcatga ctcagtccga gtcgactcgg gtcgttggtt tgctggctga ccttttgttc 360
atcaacatta tggacattgc caatgagttc aatgttccaa cttatgtata ctcccctgcc 420
ggagccggtc atcttggcct cgcgttccat ctccagacac tcaacgacaa aaagcaagat 480
gtgaccgagt tcaggaactc ggacactgag ttattggtac cgagttttgc aaacccggtt 540
cccgccgagg tcttgccgtc gatgtatgtg gataaagaag gtgggtatga ttatttgttt 600
tcattgttcc ggaggtgcag agagtcaaag gcaattatta ttaacacgtt tgaggagctg 660
gaaccctatg cgatcaattc cctccggatg gatagtatga tccctccgat ctacccggtg 720
ggacccatac taaatctcaa cggtgatggc caaaactccg atgaggctgc tgtgatcctt 780
ggttggttag atgatcaacc accttcatct gtggtgtttt tgtgctttgg tagctatgga 840
agctttcaag aaaaccaggt gaaggagatt gcaatgggtc tagagcgcag tgggcatcgc 900
ttcttgtggt ccttgcgtcc gtctatccct aaaggcgaga caaagcttca gcttaaatac 960
tcaaatttga aagaaattct cccagtagga ttcttggaca ggacatcatg cgtcggaaaa 1020
gtgattggat gggccccgca agtggccgtg ctcggacatg agtcagtcgg agggttcctg 1080
tctcattgcg gttggaattc gacattggag agtgtttggt gtggggtgcc cgttgcaaca 1140
tggccaatgt atggtgagca acaactcaat gcttttgaga tggttaagga gttaggtatt 1200
gcggtggaaa ttgaggtgga ctataagaaa gattatttta acatgaagaa tgattttatt 1260
gttagggcag aagaaatcga gacaaaaata aagaagttga tgatggatga aaataatagt 1320
gaaataagaa agaaggtaaa ggaaatgaaa gaaaagagta gggctgcaat gtctgagaat 1380
ggatcatctt ataattcatt ggcgaagcta tttgaggaaa ttatg 1425
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Met Lys Ser Glu Leu Ile Phe Leu Pro Ala Pro Ala Ile Gly His Leu
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Val Gly Met Val Glu Met Ala Lys Leu Phe Ile Ser Arg His Glu Asn
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Asp Asn Tyr Asn Lys Ser Leu Leu Thr Asn Pro Thr Pro Arg Leu Thr
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Ile Val Asn Leu Pro Glu Thr Asp Pro Gln Asn Tyr Met Leu Lys Pro
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Arg His Ala Ile Phe Pro Ser Val Ile Glu Thr Gln Lys Thr His Val
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Arg Asp Ile Ile Ser Gly Met Thr Gln Ser Glu Ser Thr Arg Val Val
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Gly Leu Leu Ala Asp Leu Leu Phe Ile Asn Ile Met Asp Ile Ala Asn
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Glu Phe Asn Val Pro Thr Tyr Val Tyr Ser Pro Ala Gly Ala Gly His
130 135 140
Leu Gly Leu Ala Phe His Leu Gln Thr Leu Asn Asp Lys Lys Gln Asp
145 150 155 160
Val Thr Glu Phe Arg Asn Ser Asp Thr Glu Leu Leu Val Pro Ser Phe
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Ala Asn Pro Val Pro Ala Glu Val Leu Pro Ser Met Tyr Val Asp Lys
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Glu Gly Gly Tyr Asp Tyr Leu Phe Ser Leu Phe Arg Arg Cys Arg Glu
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Ser Lys Ala Ile Ile Ile Asn Thr Phe Glu Glu Leu Glu Pro Tyr Ala
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Ile Asn Ser Leu Arg Met Asp Ser Met Ile Pro Pro Ile Tyr Pro Val
225 230 235 240
Gly Pro Ile Leu Asn Leu Asn Gly Asp Gly Gln Asn Ser Asp Glu Ala
245 250 255
Ala Val Ile Leu Gly Trp Leu Asp Asp Gln Pro Pro Ser Ser Val Val
260 265 270
Phe Leu Cys Phe Gly Ser Tyr Gly Ser Phe Gln Glu Asn Gln Val Lys
275 280 285
Glu Ile Ala Met Gly Leu Glu Arg Ser Gly His Arg Phe Leu Trp Ser
290 295 300
Leu Arg Pro Ser Ile Pro Lys Gly Glu Thr Lys Leu Gln Leu Lys Tyr
305 310 315 320
Ser Asn Leu Lys Glu Ile Leu Pro Val Gly Phe Leu Asp Arg Thr Ser
325 330 335
Cys Val Gly Lys Val Ile Gly Trp Ala Pro Gln Val Ala Val Leu Gly
340 345 350
His Glu Ser Val Gly Gly Phe Leu Ser His Cys Gly Trp Asn Ser Thr
355 360 365
Leu Glu Ser Val Trp Cys Gly Val Pro Val Ala Thr Trp Pro Met Tyr
370 375 380
Gly Glu Gln Gln Leu Asn Ala Phe Glu Met Val Lys Glu Leu Gly Ile
385 390 395 400
Ala Val Glu Ile Glu Val Asp Tyr Lys Lys Asp Tyr Phe Asn Met Lys
405 410 415
Asn Asp Phe Ile Val Arg Ala Glu Glu Ile Glu Thr Lys Ile Lys Lys
420 425 430
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435 440 445
Met Lys Glu Lys Ser Arg Ala Ala Met Ser Glu Asn Gly Ser Ser Tyr
450 455 460
Asn Ser Leu Ala Lys Leu Phe Glu Glu Ile Met
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<210> 3
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<220>
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atgggagcgg agctcatctt aatcccttct ccgggagttg gtcatctggt atctactgtt 60
gagattgcaa agctcctcat cagtcgagat gaacggcttt ccatcacagt ccttgtaatg 120
aagttctcgc atgacactgg tgtgactgcc tacactcggt cattgcagaa agatgctcct 180
aaccgcatag tctttgtgga ccttcctcag aacgagtctc ttatctcatc gcccaagtct 240
ttctttacta gcttcatcga gagtcagacg agtccggtta gagattctgt cagacaaatt 300
gtgagtcggt ctgattctaa taagctcgct ggcttcgtca tcgacatgtt ctgcacccca 360
atgatagacg tggcaaatga atttggagtc ccaacctatg tgttcttcac ttcaggtgct 420
gcatttcttg gcctccagtt ttaccatctg agtctcagtg atgaacataa ccaggacctt 480
accgagtata aggacacgga tgttgagtta tctatcccga gtttcatcaa cccagtgccc 540
gctaaggttt tgccttcggt gattctgaac aaggaaggat cgaccatgct ccaatctatt 600
tcccgaaggt ttaaagaagc caaggccatt ctagtcaaca cgttcgcgga gctggaacca 660
catgccatta aggcccttgg tgataactgc aagatccctc ctatctatcc cgtgggaccc 720
ataatcaacc tcaagaacaa ggagggaaca acccaaaacc atagttctga agatggtatc 780
attagctggt tggacaatca gccaccatct tcggtagtgt ttttgtgctt tgggagcttt 840
gggagctttg atgaaggcca agtcagggag atagcaaacg gattggagca gagtggacag 900
cgattcttgt ggtctctacg ccggcggcca gaaaaaatgg aattgcctaa agactatgag 960
aatcccgaag aagtgttgcc agaaggattc atagaacgaa catcagggat ggggaaggtg 1020
atcggatggg cgccacaaac ggcgattctt tcccaccctg ctgtgggagg attcgtgtct 1080
cattgtggat ggaattctac attggagagt atatggtgtg gggttccaat ggctacttgg 1140
cctatatatg cagagcagca aatcaatgcg tttgagttgg tgaaggagtt gggaatggct 1200
gtggagatca aaatggatta cagagaagat tatatttttg cacctgaaaa taatttagtt 1260
gtgaccgcag accagataga gaaaggaatg cgatgtctga tgatggatgg agagagtgaa 1320
atgaggaaga aggtggaaga gatgaaagag aagagcagaa tggccatggt gaagggcggg 1380
tcttcttaca tttcactcgg gcattttatt gaggatgtca tgcgtaatta a 1431
<210> 4
<211> 476
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT13
<400> 4
Met Gly Ala Glu Leu Ile Leu Ile Pro Ser Pro Gly Val Gly His Leu
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Val Ser Thr Val Glu Ile Ala Lys Leu Leu Ile Ser Arg Asp Glu Arg
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Leu Ser Ile Thr Val Leu Val Met Lys Phe Ser His Asp Thr Gly Val
35 40 45
Thr Ala Tyr Thr Arg Ser Leu Gln Lys Asp Ala Pro Asn Arg Ile Val
50 55 60
Phe Val Asp Leu Pro Gln Asn Glu Ser Leu Ile Ser Ser Pro Lys Ser
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Phe Phe Thr Ser Phe Ile Glu Ser Gln Thr Ser Pro Val Arg Asp Ser
85 90 95
Val Arg Gln Ile Val Ser Arg Ser Asp Ser Asn Lys Leu Ala Gly Phe
100 105 110
Val Ile Asp Met Phe Cys Thr Pro Met Ile Asp Val Ala Asn Glu Phe
115 120 125
Gly Val Pro Thr Tyr Val Phe Phe Thr Ser Gly Ala Ala Phe Leu Gly
130 135 140
Leu Gln Phe Tyr His Leu Ser Leu Ser Asp Glu His Asn Gln Asp Leu
145 150 155 160
Thr Glu Tyr Lys Asp Thr Asp Val Glu Leu Ser Ile Pro Ser Phe Ile
165 170 175
Asn Pro Val Pro Ala Lys Val Leu Pro Ser Val Ile Leu Asn Lys Glu
180 185 190
Gly Ser Thr Met Leu Gln Ser Ile Ser Arg Arg Phe Lys Glu Ala Lys
195 200 205
Ala Ile Leu Val Asn Thr Phe Ala Glu Leu Glu Pro His Ala Ile Lys
210 215 220
Ala Leu Gly Asp Asn Cys Lys Ile Pro Pro Ile Tyr Pro Val Gly Pro
225 230 235 240
Ile Ile Asn Leu Lys Asn Lys Glu Gly Thr Thr Gln Asn His Ser Ser
245 250 255
Glu Asp Gly Ile Ile Ser Trp Leu Asp Asn Gln Pro Pro Ser Ser Val
260 265 270
Val Phe Leu Cys Phe Gly Ser Phe Gly Ser Phe Asp Glu Gly Gln Val
275 280 285
Arg Glu Ile Ala Asn Gly Leu Glu Gln Ser Gly Gln Arg Phe Leu Trp
290 295 300
Ser Leu Arg Arg Arg Pro Glu Lys Met Glu Leu Pro Lys Asp Tyr Glu
305 310 315 320
Asn Pro Glu Glu Val Leu Pro Glu Gly Phe Ile Glu Arg Thr Ser Gly
325 330 335
Met Gly Lys Val Ile Gly Trp Ala Pro Gln Thr Ala Ile Leu Ser His
340 345 350
Pro Ala Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Thr Leu
355 360 365
Glu Ser Ile Trp Cys Gly Val Pro Met Ala Thr Trp Pro Ile Tyr Ala
370 375 380
Glu Gln Gln Ile Asn Ala Phe Glu Leu Val Lys Glu Leu Gly Met Ala
385 390 395 400
Val Glu Ile Lys Met Asp Tyr Arg Glu Asp Tyr Ile Phe Ala Pro Glu
405 410 415
Asn Asn Leu Val Val Thr Ala Asp Gln Ile Glu Lys Gly Met Arg Cys
420 425 430
Leu Met Met Asp Gly Glu Ser Glu Met Arg Lys Lys Val Glu Glu Met
435 440 445
Lys Glu Lys Ser Arg Met Ala Met Val Lys Gly Gly Ser Ser Tyr Ile
450 455 460
Ser Leu Gly His Phe Ile Glu Asp Val Met Arg Asn
465 470 475
<210> 5
<211> 1353
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT30
<400> 5
atggcaaccc aaaaatgttt ccgtgtcctc gtgttcccat ggttagctca tggtcacatc 60
tcccccttcc tagaactagc cagaaaactt atagaaaaag gaaacttttt catttacttc 120
tgctccactc ctataaatct catctccata aagaaaaaac ttagtggtga tgatcatcaa 180
aattacacca agtcaataca actagtggaa cacaatctac cgaccttacc tcaacttcct 240
cctcactacc acaccaccga cggcctccca ccaaatctca atcctaccct tcgaaaggca 300
tttgaaatgt caaaactatc cttccccaac accctaaaca ctttgaaacc agatctactc 360
atttgtgatg acttatttca atggccagaa atagtagctt catcacatgg tgttccggtt 420
gttcggttcc aaacatgcag cgtgacagcc gctagttttc tagctcatat ttttacgaac 480
ccagacgtta catacccttt tccagccatt tatcttcatg agtatgaaac tgatcagatc 540
aggcgttgcg ttgacgcggt ttttgaaagc ggcagagaag aatctcgcaa cctgttggtt 600
gtcaacacgt ctaaagcgat cgaggaaaaa tatttttatt attattctct acttcggggt 660
aacaccaaaa ttatgccagt tggtccgctt attcagcaag ccccaaatgg cgacgaggat 720
atgaaggtca tcgaatggct tgacaagaag gatccgtgtt caaccgtgtt cgtgtccttt 780
gggagcgagt attttatgca aaaagaagag gttgaagaga tggctcatgg tttagagctt 840
agcaacgtca atttcatctg ggtttttagg gctccggtgg gagcggaaaa ggttaagctg 900
ccgttagggt ttgttgagag ggttgggggg aggggaattg ttatggaggg gtgggcccca 960
caggcaagga ttttgggaca ttcaagtatt ggtgggtttg tgagtcattg tgggtggaat 1020
tctgtgttgg aaactataaa ttttggtgtt ccaataatag ggatgccaat gaaatttgag 1080
cagcctatga atgctaggct tctatctgaa cttggtgttt gtgttgaaat tgtgggggac 1140
gaaactagga ggtttggaag agaagaggta ggaaatgtaa taagagaagt ggttggtggg 1200
aagatagggg acgatttgag aaggaaagtg aaagaacttg gagcaaagat aaaggaaaaa 1260
caagaggaag agatggatga tgtgcttgat gaattagtac aaatttgtaa caagaaaaaa 1320
cggactgttg tacatcatca tcatcatcat tga 1353
<210> 6
<211> 450
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT30
<400> 6
Met Ala Thr Gln Lys Cys Phe Arg Val Leu Val Phe Pro Trp Leu Ala
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His Gly His Ile Ser Pro Phe Leu Glu Leu Ala Arg Lys Leu Ile Glu
20 25 30
Lys Gly Asn Phe Phe Ile Tyr Phe Cys Ser Thr Pro Ile Asn Leu Ile
35 40 45
Ser Ile Lys Lys Lys Leu Ser Gly Asp Asp His Gln Asn Tyr Thr Lys
50 55 60
Ser Ile Gln Leu Val Glu His Asn Leu Pro Thr Leu Pro Gln Leu Pro
65 70 75 80
Pro His Tyr His Thr Thr Asp Gly Leu Pro Pro Asn Leu Asn Pro Thr
85 90 95
Leu Arg Lys Ala Phe Glu Met Ser Lys Leu Ser Phe Pro Asn Thr Leu
100 105 110
Asn Thr Leu Lys Pro Asp Leu Leu Ile Cys Asp Asp Leu Phe Gln Trp
115 120 125
Pro Glu Ile Val Ala Ser Ser His Gly Val Pro Val Val Arg Phe Gln
130 135 140
Thr Cys Ser Val Thr Ala Ala Ser Phe Leu Ala His Ile Phe Thr Asn
145 150 155 160
Pro Asp Val Thr Tyr Pro Phe Pro Ala Ile Tyr Leu His Glu Tyr Glu
165 170 175
Thr Asp Gln Ile Arg Arg Cys Val Asp Ala Val Phe Glu Ser Gly Arg
180 185 190
Glu Glu Ser Arg Asn Leu Leu Val Val Asn Thr Ser Lys Ala Ile Glu
195 200 205
Glu Lys Tyr Phe Tyr Tyr Tyr Ser Leu Leu Arg Gly Asn Thr Lys Ile
210 215 220
Met Pro Val Gly Pro Leu Ile Gln Gln Ala Pro Asn Gly Asp Glu Asp
225 230 235 240
Met Lys Val Ile Glu Trp Leu Asp Lys Lys Asp Pro Cys Ser Thr Val
245 250 255
Phe Val Ser Phe Gly Ser Glu Tyr Phe Met Gln Lys Glu Glu Val Glu
260 265 270
Glu Met Ala His Gly Leu Glu Leu Ser Asn Val Asn Phe Ile Trp Val
275 280 285
Phe Arg Ala Pro Val Gly Ala Glu Lys Val Lys Leu Pro Leu Gly Phe
290 295 300
Val Glu Arg Val Gly Gly Arg Gly Ile Val Met Glu Gly Trp Ala Pro
305 310 315 320
Gln Ala Arg Ile Leu Gly His Ser Ser Ile Gly Gly Phe Val Ser His
325 330 335
Cys Gly Trp Asn Ser Val Leu Glu Thr Ile Asn Phe Gly Val Pro Ile
340 345 350
Ile Gly Met Pro Met Lys Phe Glu Gln Pro Met Asn Ala Arg Leu Leu
355 360 365
Ser Glu Leu Gly Val Cys Val Glu Ile Val Gly Asp Glu Thr Arg Arg
370 375 380
Phe Gly Arg Glu Glu Val Gly Asn Val Ile Arg Glu Val Val Gly Gly
385 390 395 400
Lys Ile Gly Asp Asp Leu Arg Arg Lys Val Lys Glu Leu Gly Ala Lys
405 410 415
Ile Lys Glu Lys Gln Glu Glu Glu Met Asp Asp Val Leu Asp Glu Leu
420 425 430
Val Gln Ile Cys Asn Lys Lys Lys Arg Thr Val Val His His His His
435 440 445
His His
450
<210> 7
<211> 26
<212> DNA
<213> 人参(Panax ginseng)
<400> 7
ccaagttcat tcaagatgaa gtcaga 26
<210> 8
<211> 28
<212> DNA
<213> oligonucleotides
<400> 8
cataatttcc tcaaatagct tcgccaat 28
<210> 9
<211> 24
<212> DNA
<213> oligonucleotides
<400> 9
actggagcag ttgcaacata caaa 24
<210> 10
<211> 24
<212> DNA
<213> oligonucleotides
<400> 10
tcatttatgc gctttcctgg atgt 24
<210> 11
<211> 21
<212> DNA
<213> oligonucleotides
<400> 11
acgagggtgg ttttaatggc a 21
<210> 12
<211> 27
<212> DNA
<213> oligonucleotides
<400> 12
acaccattca taatcataca acagtcc 27
<210> 13
<211> 29
<212> DNA
<213> oligonucleotides
<400> 13
atatggtacc atggcagtag ccggcgctg 29
<210> 14
<211> 57
<212> DNA
<213> oligonucleotides
<400> 14
gcgctctaga ttaatgatga tgatgatgat gttttattat atgttttttg gggtcgc 57
<210> 15
<211> 1428
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25-1
<400> 15
atgaagtcag aattgatatt cttgcccgcc ccggccatcg gacacctcgt gggaatggtg 60
gagatggcta aactcttcat cagtcgacat gaaaacctct cggtcaccgt cctcatcgcg 120
aaattctaca tggatacggg ggtagacaac tacaataaat cactcttaac aaaccctacc 180
ccgcgtctca caattgtaaa tctcccggaa accgaccccc aaaactatat gctcaaacca 240
cgccatgcca tctttcctag cgtcatcgag actcagaaga cacacgtgcg agacataata 300
tcaggcatga ctcagtccga gtcgactcgg gtcgttggtt tgctggctga ccttttgttc 360
atcaacatta tggacattgc caatgagttc aatgttccaa cttatgtata ctcccctgcc 420
ggagccggtc atcttggcct cgcgttccat ctccagacac tcaacgacaa aaagcaagat 480
gtgaccgagt tcaggaactc ggacactgag ttattggtac cgagttttgc aaacccggtt 540
cccgccgagg tcttgccgtc gatgtatgtg gataaagaag gtgggtatga ttatttgttt 600
tcattgttcc ggaggtgcag agagtcaaag gcaattatta ttaacacgtt tgaggagctg 660
gaaccctatg cgatcaattc cctccggatg gatagtatga tccctccgat ctacccggtg 720
ggacccatac taaatctcaa cggtgatggc caaaactccg atgaggctgc tgtgatcctt 780
ggttggttag atgatcaacc accttcatct gtggtgtttt tgtgctttgg tagctatgga 840
acctttcaag aaaaccaggt gaaggagatt gcaatgggtc tagagcgcag tgggcatcgc 900
ttcttgtggt ccttgcgtcc gtctatccct aaaggcgaga caaagcttca gcttaaatac 960
tcaaatttgg aagaaattct cccagtcgga ttcttggaca ggacatcatg cgtcggaaaa 1020
gttattggat gggccccgca agtggcggtg ctcggacacg aggcagtcgg agggttcctg 1080
tctcattgtg gttggaattc gacattagag agtgtgtggt gtggcgtgcc cgtcgcaaca 1140
tggccaatgt acggcgagca acaactcaat gcttttgaga tggttaagga gttaggtatt 1200
gcggtggaaa ttgaggtgga ctataagaat gaatatttta acatgaataa tgattttatt 1260
gttagggcag aagaaatcga gacgaaaata aagaagttga tgatggatga aaagaatagt 1320
gaaataagga agaaggtaaa ggaaatgaaa gaaaagagta ggcttgcaat gtctgagaat 1380
ggatcatctt ataattcatt ggcgaagcta tttgaggaaa tcatgtaa 1428
<210> 16
<211> 475
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25-1
<400> 16
Met Lys Ser Glu Leu Ile Phe Leu Pro Ala Pro Ala Ile Gly His Leu
1 5 10 15
Val Gly Met Val Glu Met Ala Lys Leu Phe Ile Ser Arg His Glu Asn
20 25 30
Leu Ser Val Thr Val Leu Ile Ala Lys Phe Tyr Met Asp Thr Gly Val
35 40 45
Asp Asn Tyr Asn Lys Ser Leu Leu Thr Asn Pro Thr Pro Arg Leu Thr
50 55 60
Ile Val Asn Leu Pro Glu Thr Asp Pro Gln Asn Tyr Met Leu Lys Pro
65 70 75 80
Arg His Ala Ile Phe Pro Ser Val Ile Glu Thr Gln Lys Thr His Val
85 90 95
Arg Asp Ile Ile Ser Gly Met Thr Gln Ser Glu Ser Thr Arg Val Val
100 105 110
Gly Leu Leu Ala Asp Leu Leu Phe Ile Asn Ile Met Asp Ile Ala Asn
115 120 125
Glu Phe Asn Val Pro Thr Tyr Val Tyr Ser Pro Ala Gly Ala Gly His
130 135 140
Leu Gly Leu Ala Phe His Leu Gln Thr Leu Asn Asp Lys Lys Gln Asp
145 150 155 160
Val Thr Glu Phe Arg Asn Ser Asp Thr Glu Leu Leu Val Pro Ser Phe
165 170 175
Ala Asn Pro Val Pro Ala Glu Val Leu Pro Ser Met Tyr Val Asp Lys
180 185 190
Glu Gly Gly Tyr Asp Tyr Leu Phe Ser Leu Phe Arg Arg Cys Arg Glu
195 200 205
Ser Lys Ala Ile Ile Ile Asn Thr Phe Glu Glu Leu Glu Pro Tyr Ala
210 215 220
Ile Asn Ser Leu Arg Met Asp Ser Met Ile Pro Pro Ile Tyr Pro Val
225 230 235 240
Gly Pro Ile Leu Asn Leu Asn Gly Asp Gly Gln Asn Ser Asp Glu Ala
245 250 255
Ala Val Ile Leu Gly Trp Leu Asp Asp Gln Pro Pro Ser Ser Val Val
260 265 270
Phe Leu Cys Phe Gly Ser Tyr Gly Thr Phe Gln Glu Asn Gln Val Lys
275 280 285
Glu Ile Ala Met Gly Leu Glu Arg Ser Gly His Arg Phe Leu Trp Ser
290 295 300
Leu Arg Pro Ser Ile Pro Lys Gly Glu Thr Lys Leu Gln Leu Lys Tyr
305 310 315 320
Ser Asn Leu Glu Glu Ile Leu Pro Val Gly Phe Leu Asp Arg Thr Ser
325 330 335
Cys Val Gly Lys Val Ile Gly Trp Ala Pro Gln Val Ala Val Leu Gly
340 345 350
His Glu Ala Val Gly Gly Phe Leu Ser His Cys Gly Trp Asn Ser Thr
355 360 365
Leu Glu Ser Val Trp Cys Gly Val Pro Val Ala Thr Trp Pro Met Tyr
370 375 380
Gly Glu Gln Gln Leu Asn Ala Phe Glu Met Val Lys Glu Leu Gly Ile
385 390 395 400
Ala Val Glu Ile Glu Val Asp Tyr Lys Asn Glu Tyr Phe Asn Met Asn
405 410 415
Asn Asp Phe Ile Val Arg Ala Glu Glu Ile Glu Thr Lys Ile Lys Lys
420 425 430
Leu Met Met Asp Glu Lys Asn Ser Glu Ile Arg Lys Lys Val Lys Glu
435 440 445
Met Lys Glu Lys Ser Arg Leu Ala Met Ser Glu Asn Gly Ser Ser Tyr
450 455 460
Asn Ser Leu Ala Lys Leu Phe Glu Glu Ile Met
465 470 475
<210> 17
<211> 1428
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25-3
<400> 17
atgaagtcag aattgatatt cttgcccgcc ccggccatcg gacacctcgt gggaatggtg 60
gagatggcta aactcttcat cagtcgacat gaaaatctct cggtcaccgt cctcatcgcg 120
aaattctaca tggatacggg ggtagacaac tacaataaat cactcttaac aaaccctacc 180
ccgcgtctca caattgtaaa tctcccggaa accgaccccc aaaactatat gctcaaacca 240
cgccacgcca tctttcctag cgtcatcgag actcagaaga cacacgtgcg agacataata 300
tccggtatga ctcagtccga gtcgactcag gtcgttggtt tgctggctga ccttttgttc 360
atcaacatca tggacattgc caatgagttc aatgttccaa cttatgtata ctcccctgcc 420
ggagccggtc atcttggcct cgcgttccat ctccagacac tcaacgacaa aaaacaagat 480
gtgaccgagt tcaggaactc ggatactgag ttattggtac cgagttttgc aaacccggtt 540
cccgccgagg tcttgccgtc gatgtatgtg gataaagaag gtgggtatga ttatctgttt 600
tcattgttcc ggaggtgcag agagtcaaag gcaattatta ttaacacgtt tgaggagctg 660
gaaccctatg cgatcaattc cctccggatg gatagtatga tccctccgat ctacccggtg 720
ggacccatac taaatctcaa cggtgatggc caaaactccg atgaggctgc tgtgatcctt 780
ggttggttag atgatcaacc accttcatct gtggtgtttt tgtgctttgg tagctatgga 840
agctttcaag aaaaccaggt gaaggagatt gcaatgggtc tagagcgcag tgggcatcgc 900
ttcttgtggt ccttgcgtcc gtctatccct aaaggcgaga caaagcttca gcttaaatac 960
tcaaatttga aagaaattct cccagtagga ttcttggaca ggacatcatg cgtcggaaaa 1020
gtgattggat gggccccgca agtggccgtg ctcggacatg agtcagtcgg agggttcctg 1080
tctcattgcg gttggaattc gacattggag agtgtttggt gtggggtgcc cgttgcaaca 1140
tggccaatgt atggtgagca acaactcaat gcttttgaga tggttaagga gttaggtatt 1200
gcggtggaaa ttgaggtgga ctataagaaa gattatttta acatgaagaa tgattttatt 1260
gttagggcag aagaaatcga gacaaaaata aagaagttga tgatggatga aaataatagt 1320
gaaataagaa agaaggtaaa ggaaatgaaa gaaaagagta gggctgcaat gtctgagaat 1380
ggatcatctt ataattcatt ggcgaagcta tttgaggaaa ttatgtaa 1428
<210> 18
<211> 475
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25-3
<400> 18
Met Lys Ser Glu Leu Ile Phe Leu Pro Ala Pro Ala Ile Gly His Leu
1 5 10 15
Val Gly Met Val Glu Met Ala Lys Leu Phe Ile Ser Arg His Glu Asn
20 25 30
Leu Ser Val Thr Val Leu Ile Ala Lys Phe Tyr Met Asp Thr Gly Val
35 40 45
Asp Asn Tyr Asn Lys Ser Leu Leu Thr Asn Pro Thr Pro Arg Leu Thr
50 55 60
Ile Val Asn Leu Pro Glu Thr Asp Pro Gln Asn Tyr Met Leu Lys Pro
65 70 75 80
Arg His Ala Ile Phe Pro Ser Val Ile Glu Thr Gln Lys Thr His Val
85 90 95
Arg Asp Ile Ile Ser Gly Met Thr Gln Ser Glu Ser Thr Gln Val Val
100 105 110
Gly Leu Leu Ala Asp Leu Leu Phe Ile Asn Ile Met Asp Ile Ala Asn
115 120 125
Glu Phe Asn Val Pro Thr Tyr Val Tyr Ser Pro Ala Gly Ala Gly His
130 135 140
Leu Gly Leu Ala Phe His Leu Gln Thr Leu Asn Asp Lys Lys Gln Asp
145 150 155 160
Val Thr Glu Phe Arg Asn Ser Asp Thr Glu Leu Leu Val Pro Ser Phe
165 170 175
Ala Asn Pro Val Pro Ala Glu Val Leu Pro Ser Met Tyr Val Asp Lys
180 185 190
Glu Gly Gly Tyr Asp Tyr Leu Phe Ser Leu Phe Arg Arg Cys Arg Glu
195 200 205
Ser Lys Ala Ile Ile Ile Asn Thr Phe Glu Glu Leu Glu Pro Tyr Ala
210 215 220
Ile Asn Ser Leu Arg Met Asp Ser Met Ile Pro Pro Ile Tyr Pro Val
225 230 235 240
Gly Pro Ile Leu Asn Leu Asn Gly Asp Gly Gln Asn Ser Asp Glu Ala
245 250 255
Ala Val Ile Leu Gly Trp Leu Asp Asp Gln Pro Pro Ser Ser Val Val
260 265 270
Phe Leu Cys Phe Gly Ser Tyr Gly Ser Phe Gln Glu Asn Gln Val Lys
275 280 285
Glu Ile Ala Met Gly Leu Glu Arg Ser Gly His Arg Phe Leu Trp Ser
290 295 300
Leu Arg Pro Ser Ile Pro Lys Gly Glu Thr Lys Leu Gln Leu Lys Tyr
305 310 315 320
Ser Asn Leu Lys Glu Ile Leu Pro Val Gly Phe Leu Asp Arg Thr Ser
325 330 335
Cys Val Gly Lys Val Ile Gly Trp Ala Pro Gln Val Ala Val Leu Gly
340 345 350
His Glu Ser Val Gly Gly Phe Leu Ser His Cys Gly Trp Asn Ser Thr
355 360 365
Leu Glu Ser Val Trp Cys Gly Val Pro Val Ala Thr Trp Pro Met Tyr
370 375 380
Gly Glu Gln Gln Leu Asn Ala Phe Glu Met Val Lys Glu Leu Gly Ile
385 390 395 400
Ala Val Glu Ile Glu Val Asp Tyr Lys Lys Asp Tyr Phe Asn Met Lys
405 410 415
Asn Asp Phe Ile Val Arg Ala Glu Glu Ile Glu Thr Lys Ile Lys Lys
420 425 430
Leu Met Met Asp Glu Asn Asn Ser Glu Ile Arg Lys Lys Val Lys Glu
435 440 445
Met Lys Glu Lys Ser Arg Ala Ala Met Ser Glu Asn Gly Ser Ser Tyr
450 455 460
Asn Ser Leu Ala Lys Leu Phe Glu Glu Ile Met
465 470 475
<210> 19
<211> 1419
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25-5
<400> 19
atgaagtcag aattgatatt cgtgcccgtc ccggccttcg ggcacctcgt ggggatggtg 60
gagatggcta aacttttcat cagtcgacac gaaaacctat cagtcaccgt cctcatttcg 120
aaatttttca ttgatacggg gatagacaac tacaataaat cactcttagc gaaacctacc 180
ccgcgtctca caattataaa tctcccggaa atcgatcccc aaaaatattt gctcaaacca 240
cgttgcgcca tctttccttc cctcatcgag aatcagaaga cacacgtgcg agacgtaatg 300
tcccgcatga ctcagtccga gtcgactcgg gtcgttggtt tgctggcaga cattttgttc 360
gtcgacatct tcgacattgc cgatgagttc aatgttccaa cttatgtata ctcccctgcc 420
ggagccggtt ttcttggcct cgcgttccat ctccagacac tcaacgacga caaaaagcaa 480
gatgtgaccg agttcaggaa ttcggacact gagttattgg taccgagttt tgcaaacccg 540
gtccccgccg agttcttgcc gtcgatattt ttggaaaaag atggtaggca tgatgttttg 600
ttatcattgt actggaggtg cagggaggca aagggaatta ttgttaacac gtttgaggag 660
ctggaaccct atgcgatcaa ttccctccgg atggatagta tgatccctcc gatctacccg 720
gtgggaccca tactaaatct caacggtgag ggacaaaact ccgatgaggc tgctgtgatc 780
cttggttggt tagatgatca accaccttca tctgtggtgt ttttgtgctt tggtagcttt 840
ggaagctttc cagaaaacca ggtgaaggag attgcaatgg gtttagagcg cagcgggcat 900
cgcttcttgt ggtccttgcg tccgtgtatc tctgaaggtg agacaacgct tcaacttaaa 960
tactcaaatt tggaacttcc ggccggattc ttggatagga catcatgcgt cggaaaagtg 1020
attggatggg ccccacaaat ggccatccta gcacacgagg cagtcggagg gttcgtgtct 1080
cattgtggtt ggaattcggt actagagagt gtgtggtatg gcatgcctgt cgcaacatgg 1140
ccaatgtacg gtgagcaaca actcaacgct tttgagatgg ttaaggagtt gggtcttgcg 1200
gtggaaattg aggtggacta taggaatgaa tataacaagt ctgattttat tgttaaggct 1260
gacgaaattg agacaaaaat aaagaagttg atgatggatg gaaagaatag taaaataagg 1320
aagaaggtaa aggaaatgaa agaaaagagt agggttgcca tgtcggagaa tgggtcatct 1380
tatacttcat tggcgaagct atttgaggaa attatgtaa 1419
<210> 20
<211> 472
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT25-5
<400> 20
Met Lys Ser Glu Leu Ile Phe Val Pro Val Pro Ala Phe Gly His Leu
1 5 10 15
Val Gly Met Val Glu Met Ala Lys Leu Phe Ile Ser Arg His Glu Asn
20 25 30
Leu Ser Val Thr Val Leu Ile Ser Lys Phe Phe Ile Asp Thr Gly Ile
35 40 45
Asp Asn Tyr Asn Lys Ser Leu Leu Ala Lys Pro Thr Pro Arg Leu Thr
50 55 60
Ile Ile Asn Leu Pro Glu Ile Asp Pro Gln Lys Tyr Leu Leu Lys Pro
65 70 75 80
Arg Cys Ala Ile Phe Pro Ser Leu Ile Glu Asn Gln Lys Thr His Val
85 90 95
Arg Asp Val Met Ser Arg Met Thr Gln Ser Glu Ser Thr Arg Val Val
100 105 110
Gly Leu Leu Ala Asp Ile Leu Phe Val Asp Ile Phe Asp Ile Ala Asp
115 120 125
Glu Phe Asn Val Pro Thr Tyr Val Tyr Ser Pro Ala Gly Ala Gly Phe
130 135 140
Leu Gly Leu Ala Phe His Leu Gln Thr Leu Asn Asp Asp Lys Lys Gln
145 150 155 160
Asp Val Thr Glu Phe Arg Asn Ser Asp Thr Glu Leu Leu Val Pro Ser
165 170 175
Phe Ala Asn Pro Val Pro Ala Glu Phe Leu Pro Ser Ile Phe Leu Glu
180 185 190
Lys Asp Gly Arg His Asp Val Leu Leu Ser Leu Tyr Trp Arg Cys Arg
195 200 205
Glu Ala Lys Gly Ile Ile Val Asn Thr Phe Glu Glu Leu Glu Pro Tyr
210 215 220
Ala Ile Asn Ser Leu Arg Met Asp Ser Met Ile Pro Pro Ile Tyr Pro
225 230 235 240
Val Gly Pro Ile Leu Asn Leu Asn Gly Glu Gly Gln Asn Ser Asp Glu
245 250 255
Ala Ala Val Ile Leu Gly Trp Leu Asp Asp Gln Pro Pro Ser Ser Val
260 265 270
Val Phe Leu Cys Phe Gly Ser Phe Gly Ser Phe Pro Glu Asn Gln Val
275 280 285
Lys Glu Ile Ala Met Gly Leu Glu Arg Ser Gly His Arg Phe Leu Trp
290 295 300
Ser Leu Arg Pro Cys Ile Ser Glu Gly Glu Thr Thr Leu Gln Leu Lys
305 310 315 320
Tyr Ser Asn Leu Glu Leu Pro Ala Gly Phe Leu Asp Arg Thr Ser Cys
325 330 335
Val Gly Lys Val Ile Gly Trp Ala Pro Gln Met Ala Ile Leu Ala His
340 345 350
Glu Ala Val Gly Gly Phe Val Ser His Cys Gly Trp Asn Ser Val Leu
355 360 365
Glu Ser Val Trp Tyr Gly Met Pro Val Ala Thr Trp Pro Met Tyr Gly
370 375 380
Glu Gln Gln Leu Asn Ala Phe Glu Met Val Lys Glu Leu Gly Leu Ala
385 390 395 400
Val Glu Ile Glu Val Asp Tyr Arg Asn Glu Tyr Asn Lys Ser Asp Phe
405 410 415
Ile Val Lys Ala Asp Glu Ile Glu Thr Lys Ile Lys Lys Leu Met Met
420 425 430
Asp Gly Lys Asn Ser Lys Ile Arg Lys Lys Val Lys Glu Met Lys Glu
435 440 445
Lys Ser Arg Val Ala Met Ser Glu Asn Gly Ser Ser Tyr Thr Ser Leu
450 455 460
Ala Lys Leu Phe Glu Glu Ile Met
465 470
<210> 21
<211> 1488
<212> DNA
<213> 欧洲山芥(Barbarea vulgaris)
<220>
<221> misc_feature
<223> 3GT1
<400> 21
atggtttccg aaatcaccca taaatcttat cctcttcact ttgttctctt ccctttcatg 60
gctcaaggcc acatgattcc catggttgat attgcaaggc tcttggctca gcgcggtgtg 120
aaaataacaa ttgtcacaac gccgcacaat gcagcgaggt tcgagaatgt cctaagccgt 180
gccattgagt ctggcttgcc catcagcata gtgcaagtca agcttccatc tcaagaagct 240
ggcttaccag aaggaaatga gactttcgat tcacttgtct caacaaagtt gctggtacct 300
ttctttaaag cggttaacat gcttgaagaa ccggtccaga agctctttga agagatgagc 360
cctcaaccaa gctgtataat ttctgatttt tgtttgcctt atacaagcaa aatcgccaag 420
aagttcaata tcccaaagat cctcttccat ggcatgtgtt gcttttgtct tctgtgtatg 480
catgttttac gcaagaaccg tgagatcttg gaaaacttaa agtctgacaa ggagcatttc 540
gttgttcctt attttcctga tcgagttgaa ttcacaagac ctcaagttcc attggcaaca 600
tatgttcctg gggaatggca cgagatcaag gaggatatgg tagaagcgga taagacttcc 660
tatggtgtga tagtcaacac atatcaagag ctcgagcctg cttatgccaa cggctacaag 720
gaggcaaggt ctggtaaagc atggaccatt ggacctgttt ccttgtgcaa caaggtggga 780
gccgacaaag cagagagggg aaacaaagca gacattgatc aagatgagtg tcttaaatgg 840
cttgattcta aagaagaagg ttcggttcta tatgtttgcc ttggaagtat ctgcagtctt 900
cctctgtctc agctcaagga gctggggcta ggccttgagg aatcccaaag acctttcatt 960
tgggtcgtaa gaggttggga gaagaacaaa gagttacttg agtggttctc ggagagcgga 1020
tttgaagaaa gagtaaaaga cagagggctt ctcatcaaag gatggtcacc tcaaatgctt 1080
atccttgcac atcattccgt tggagggttc ttaacacact gtggatggaa ctcgaccctc 1140
gaaggaatca cttcaggcgt tccattgctc acttggccac tgtttggaga ccaattctgc 1200
aaccaaaaac ttgtcgtgca ggtgctaaaa gtgggtgtaa gtgccggggt tgaagaggtt 1260
acgaattggg gagaagagga gaaaatagga gtattagtgg ataaagaggg agtgaagaag 1320
gcagtggaag aattaatggg tgagagtgat gatgctaaag aaataagaaa aagagtcaaa 1380
gagcttggac aattagctca caaggctgtg gaggaaggag gctcatctca ttctaatatc 1440
acatccttgc tagaagacat aatgcaacta gcacaaccta ataattaa 1488
<210> 22
<211> 495
<212> PRT
<213> 欧洲山芥(Barbarea vulgaris)
<220>
<221> misc_feature
<223> 3GT1
<400> 22
Met Val Ser Glu Ile Thr His Lys Ser Tyr Pro Leu His Phe Val Leu
1 5 10 15
Phe Pro Phe Met Ala Gln Gly His Met Ile Pro Met Val Asp Ile Ala
20 25 30
Arg Leu Leu Ala Gln Arg Gly Val Lys Ile Thr Ile Val Thr Thr Pro
35 40 45
His Asn Ala Ala Arg Phe Glu Asn Val Leu Ser Arg Ala Ile Glu Ser
50 55 60
Gly Leu Pro Ile Ser Ile Val Gln Val Lys Leu Pro Ser Gln Glu Ala
65 70 75 80
Gly Leu Pro Glu Gly Asn Glu Thr Phe Asp Ser Leu Val Ser Thr Lys
85 90 95
Leu Leu Val Pro Phe Phe Lys Ala Val Asn Met Leu Glu Glu Pro Val
100 105 110
Gln Lys Leu Phe Glu Glu Met Ser Pro Gln Pro Ser Cys Ile Ile Ser
115 120 125
Asp Phe Cys Leu Pro Tyr Thr Ser Lys Ile Ala Lys Lys Phe Asn Ile
130 135 140
Pro Lys Ile Leu Phe His Gly Met Cys Cys Phe Cys Leu Leu Cys Met
145 150 155 160
His Val Leu Arg Lys Asn Arg Glu Ile Leu Glu Asn Leu Lys Ser Asp
165 170 175
Lys Glu His Phe Val Val Pro Tyr Phe Pro Asp Arg Val Glu Phe Thr
180 185 190
Arg Pro Gln Val Pro Leu Ala Thr Tyr Val Pro Gly Glu Trp His Glu
195 200 205
Ile Lys Glu Asp Met Val Glu Ala Asp Lys Thr Ser Tyr Gly Val Ile
210 215 220
Val Asn Thr Tyr Gln Glu Leu Glu Pro Ala Tyr Ala Asn Gly Tyr Lys
225 230 235 240
Glu Ala Arg Ser Gly Lys Ala Trp Thr Ile Gly Pro Val Ser Leu Cys
245 250 255
Asn Lys Val Gly Ala Asp Lys Ala Glu Arg Gly Asn Lys Ala Asp Ile
260 265 270
Asp Gln Asp Glu Cys Leu Lys Trp Leu Asp Ser Lys Glu Glu Gly Ser
275 280 285
Val Leu Tyr Val Cys Leu Gly Ser Ile Cys Ser Leu Pro Leu Ser Gln
290 295 300
Leu Lys Glu Leu Gly Leu Gly Leu Glu Glu Ser Gln Arg Pro Phe Ile
305 310 315 320
Trp Val Val Arg Gly Trp Glu Lys Asn Lys Glu Leu Leu Glu Trp Phe
325 330 335
Ser Glu Ser Gly Phe Glu Glu Arg Val Lys Asp Arg Gly Leu Leu Ile
340 345 350
Lys Gly Trp Ser Pro Gln Met Leu Ile Leu Ala His His Ser Val Gly
355 360 365
Gly Phe Leu Thr His Cys Gly Trp Asn Ser Thr Leu Glu Gly Ile Thr
370 375 380
Ser Gly Val Pro Leu Leu Thr Trp Pro Leu Phe Gly Asp Gln Phe Cys
385 390 395 400
Asn Gln Lys Leu Val Val Gln Val Leu Lys Val Gly Val Ser Ala Gly
405 410 415
Val Glu Glu Val Thr Asn Trp Gly Glu Glu Glu Lys Ile Gly Val Leu
420 425 430
Val Asp Lys Glu Gly Val Lys Lys Ala Val Glu Glu Leu Met Gly Glu
435 440 445
Ser Asp Asp Ala Lys Glu Ile Arg Lys Arg Val Lys Glu Leu Gly Gln
450 455 460
Leu Ala His Lys Ala Val Glu Glu Gly Gly Ser Ser His Ser Asn Ile
465 470 475 480
Thr Ser Leu Leu Glu Asp Ile Met Gln Leu Ala Gln Pro Asn Asn
485 490 495
<210> 23
<211> 1488
<212> DNA
<213> 欧洲山芥(Barbarea vulgaris)
<220>
<221> misc_feature
<223> 3GT2
<400> 23
atggtttccg aaatcaccca taaatcttat cctcttcact ttgttctctt ccctttcatg 60
gctcaaggcc acatgattcc catggttgat attgcaaggc tcttggccca gcgcggtgtg 120
aaaataacaa ttgtcacaac cccgcacaat gcagcgaggt tcaagaatgt cctaagtcgt 180
gccattgagt ctggcttgcc catcagcata gtgcaagtca agcttccatc tcaagaagct 240
ggcttaccag aaggaaatga gactctcgat tcacttgtct cgatggagtt gatgatacat 300
ttcttaaaag cggttaacat gctggaagaa ccggtccaga agctctttga agagatgagc 360
cctcaaccaa gctgtataat ttctgatttt tgtttgcctt atacaagcaa aatcgccaag 420
aagttcaata tcccaaagat cctcttccat ggcatgtgct gcttttgtct tctgtgtatg 480
catattttac gcaagaaccg tgagatcgtg gaaaacttaa agtctgacaa ggagcatttc 540
gttgttcctt attttcctga tcgagttgaa ttcacaagac ctcaagttcc agtggcaaca 600
tatgttcctg gagactggca cgagatcacg gaggatatgg tagaagcgga taagacttcc 660
tatggtgtga tagtcaacac atatcaagag ctcgagcctg cttatgccaa cgactacaag 720
gaggcaaggt ctggtaaagc atggaccatt ggacctgttt ccttgtgcaa caaggtggga 780
gcggacaaag cagagagggg aaacaaagca gacattgatc aagatgagtg tcttaaatgg 840
cttaattcta aagaagaagg ttcggttcta tatgtttgcc ttggaagtat ctgcaatctt 900
cctctgtctc agctcaagga gctcgggcta ggccttgagg aatcccaaag acctttcatt 960
tgggtcataa gaggttggga gaagaacaaa gagttacatg agtggttctc ggagagcgga 1020
ttcgaagaaa gaatcaaaga cagaggactt ctcatcaaag gatgggctcc tcaaatgctt 1080
atactttcac atcattccgt tggagggttc ttaacacact gtggatggaa ctcgactctt 1140
gaggggctaa ccgctggtct accactgctg acatggccgc ttttcgcaga ccagttctgc 1200
aacgagaaac ttgccgtgca ggtattaaaa gccggtgtaa gcgccggggt tgaccagcct 1260
atgaaatggg gagaagagga gaaaatagga gtgttggtgg ataaagaagg agtgaagaag 1320
gcagtggaag aattaatggg tgagagtgat gatgctaaag agataagaag aagagccaaa 1380
gagcttggag aattagctca caaggctgtg gaggaaggag gctcatctca ttctaatatc 1440
acatcccttc tagaagacat aatgcaacta gcacaatcca ataattaa 1488
<210> 24
<211> 495
<212> PRT
<213> 欧洲山芥(Barbarea vulgaris)
<220>
<221> misc_feature
<223> 3GT2
<400> 24
Met Val Ser Glu Ile Thr His Lys Ser Tyr Pro Leu His Phe Val Leu
1 5 10 15
Phe Pro Phe Met Ala Gln Gly His Met Ile Pro Met Val Asp Ile Ala
20 25 30
Arg Leu Leu Ala Gln Arg Gly Val Lys Ile Thr Ile Val Thr Thr Pro
35 40 45
His Asn Ala Ala Arg Phe Lys Asn Val Leu Ser Arg Ala Ile Glu Ser
50 55 60
Gly Leu Pro Ile Ser Ile Val Gln Val Lys Leu Pro Ser Gln Glu Ala
65 70 75 80
Gly Leu Pro Glu Gly Asn Glu Thr Leu Asp Ser Leu Val Ser Met Glu
85 90 95
Leu Met Ile His Phe Leu Lys Ala Val Asn Met Leu Glu Glu Pro Val
100 105 110
Gln Lys Leu Phe Glu Glu Met Ser Pro Gln Pro Ser Cys Ile Ile Ser
115 120 125
Asp Phe Cys Leu Pro Tyr Thr Ser Lys Ile Ala Lys Lys Phe Asn Ile
130 135 140
Pro Lys Ile Leu Phe His Gly Met Cys Cys Phe Cys Leu Leu Cys Met
145 150 155 160
His Ile Leu Arg Lys Asn Arg Glu Ile Val Glu Asn Leu Lys Ser Asp
165 170 175
Lys Glu His Phe Val Val Pro Tyr Phe Pro Asp Arg Val Glu Phe Thr
180 185 190
Arg Pro Gln Val Pro Val Ala Thr Tyr Val Pro Gly Asp Trp His Glu
195 200 205
Ile Thr Glu Asp Met Val Glu Ala Asp Lys Thr Ser Tyr Gly Val Ile
210 215 220
Val Asn Thr Tyr Gln Glu Leu Glu Pro Ala Tyr Ala Asn Asp Tyr Lys
225 230 235 240
Glu Ala Arg Ser Gly Lys Ala Trp Thr Ile Gly Pro Val Ser Leu Cys
245 250 255
Asn Lys Val Gly Ala Asp Lys Ala Glu Arg Gly Asn Lys Ala Asp Ile
260 265 270
Asp Gln Asp Glu Cys Leu Lys Trp Leu Asn Ser Lys Glu Glu Gly Ser
275 280 285
Val Leu Tyr Val Cys Leu Gly Ser Ile Cys Asn Leu Pro Leu Ser Gln
290 295 300
Leu Lys Glu Leu Gly Leu Gly Leu Glu Glu Ser Gln Arg Pro Phe Ile
305 310 315 320
Trp Val Ile Arg Gly Trp Glu Lys Asn Lys Glu Leu His Glu Trp Phe
325 330 335
Ser Glu Ser Gly Phe Glu Glu Arg Ile Lys Asp Arg Gly Leu Leu Ile
340 345 350
Lys Gly Trp Ala Pro Gln Met Leu Ile Leu Ser His His Ser Val Gly
355 360 365
Gly Phe Leu Thr His Cys Gly Trp Asn Ser Thr Leu Glu Gly Leu Thr
370 375 380
Ala Gly Leu Pro Leu Leu Thr Trp Pro Leu Phe Ala Asp Gln Phe Cys
385 390 395 400
Asn Glu Lys Leu Ala Val Gln Val Leu Lys Ala Gly Val Ser Ala Gly
405 410 415
Val Asp Gln Pro Met Lys Trp Gly Glu Glu Glu Lys Ile Gly Val Leu
420 425 430
Val Asp Lys Glu Gly Val Lys Lys Ala Val Glu Glu Leu Met Gly Glu
435 440 445
Ser Asp Asp Ala Lys Glu Ile Arg Arg Arg Ala Lys Glu Leu Gly Glu
450 455 460
Leu Ala His Lys Ala Val Glu Glu Gly Gly Ser Ser His Ser Asn Ile
465 470 475 480
Thr Ser Leu Leu Glu Asp Ile Met Gln Leu Ala Gln Ser Asn Asn
485 490 495
<210> 25
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29
<400> 25
atggataacc aaaaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcgaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aaggataagg attcctctgc ttctataaaa 180
ctagttgagc ttcatcttcc atcttcccct gatcttcctc ctcactatca caccacaaat 240
ggcctccctt cccatctcat gctcccactc agaaacgcct ttgaaactgc aggccccacc 300
ttctctgaaa tccttaaaac cttaaacccc gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttatttcct aaccacggca 420
gcagccagct cttccattgg cctacatgct ttcaaaaacc caggtgaaaa atacccattt 480
ccagattttt atgataacag taatattacc cctgaaccac cttctgcaga taacatgaag 540
ctacttcatg attttatcgc ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttagagaac tagaagggaa atatattgat ttgctttcca ctttatctga taaaactttg 660
gttcctgttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagattataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
acggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg gattttacca 900
gaggggtttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attgcggaga gtatgaagtt tggggttcca gtaattgcca tggccaggca tcttgatcag 1080
cctttgaatg gtaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgag 1140
aatgggaagt ataagagaga agggattgca gaggtaataa gaaaagtggt tgtggagaaa 1200
agtggggagg ttatcaggag gaaagcaagg gagttgagtg agaaaatgaa agagaaagga 1260
gagcaagaga ttgatagggc attggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gaacaatag 1329
<210> 26
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29
<400> 26
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asp Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Leu Pro Leu Arg Asn Ala Phe Glu Thr
85 90 95
Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala Ala Ala Ser Ser
130 135 140
Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Pro Asp Phe Tyr Asp Asn Ser Asn Ile Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu Glu Gly Lys Tyr
195 200 205
Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Val Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Thr Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Ile Leu Pro Glu Gly Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly Val Pro Val Ile
340 345 350
Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Ala Leu Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440
<210> 27
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-3
<400> 27
atggataacc aagaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcaaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aagaataagg attcctctgc ttctataaaa 180
ctagttgagc ttcatcttcc atcttcccct gatcttcctc ctcactatca caccacaaat 240
ggcctccctt cccatctcat ggtcccactc ataaacgcct ttgaaacagc aggccccacc 300
ttctctgaaa tccttaaaac cttaaacccc gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttatttcct aaccacggca 420
gcagccagct cttccattgg cctacatgct ttcaaaaacc caggtgaaaa atacccattt 480
ccagattttt atgataacag taataatacc cctgaaccac cttctgcaga taacatgaag 540
ctacttcatg attttatcgc ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttatagaac tagaagggaa atatatcgat ttgctttcca ctttatctga taaaactttg 660
gttcctgttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagattataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
atggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg ggttttacca 900
gagggatttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attgcggaga gtatgaagtt tggggttcca gtaattgcca tggccaggca tcttgatcag 1080
cctttgaatg gtaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgaa 1140
aatgggaagt ataagagaga agggattgca gaggtaataa gaaaagtcgt tgtggagaaa 1200
agtggggagg ttatgaggag gaaagcaagg gaattgagtg agaaaatgaa agagaaagga 1260
gaggaagaga ttgatagggc agtggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gcacaatag 1329
<210> 28
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-3
<400> 28
Met Asp Asn Gln Glu Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Val Pro Leu Ile Asn Ala Phe Glu Thr
85 90 95
Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala Ala Ala Ser Ser
130 135 140
Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Pro Asp Phe Tyr Asp Asn Ser Asn Asn Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Ile Glu Leu Glu Gly Lys Tyr
195 200 205
Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Val Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro Glu Gly Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly Val Pro Val Ile
340 345 350
Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Met Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Lys Gly Glu Glu Glu Ile Asp Arg Ala Val Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Ala Gln
435 440
<210> 29
<211> 20
<212> DNA
<213> oligonucleotides
<400> 29
atggtttccg aaatcaccca 20
<210> 30
<211> 24
<212> DNA
<213> oligonucleotides
<400> 30
ttaattattr grttgtgcta gttg 24
<210> 31
<211> 35
<212> DNA
<213> oligonucleotides
<400> 31
gactggatcc atggtttccg aaatcaccca taaat 35
<210> 32
<211> 30
<212> DNA
<213> oligonucleotides
<400> 32
gcgcgtcgac attattaggt tgtgctagtt 30
<210> 33
<211> 30
<212> DNA
<213> oligonucleotides
<400> 33
gcgcgtcgac attattggat tgtgctagtt 30
<210> 34
<211> 26
<212> DNA
<213> oligonucleotides
<400> 34
agtaagaaaa acagagttca tcatgg 26
<210> 35
<211> 20
<212> DNA
<213> oligonucleotides
<400> 35
gcctcggtta ggctagctgt 20
<210> 36
<211> 35
<212> DNA
<213> oligonucleotides
<400> 36
ggcgggtacc atggataacc aaaaaggtag aatca 35
<210> 37
<211> 56
<212> DNA
<213> oligonucleotides
<400> 37
gccgctcgag tcaatgatga tgatgatgat gttgttcatc tttcttcttc ttacaa 56
<210> 38
<211> 35
<212> DNA
<213> oligonucleotides
<400> 38
ggcgggtacc atggataacc aagaaggtag aatca 35
<210> 39
<211> 56
<212> DNA
<213> oligonucleotides
<400> 39
gccgctcgag tcaatgatga tgatgatgat gttgtgcatc tttcttcttc ttacaa 56
<210> 40
<211> 1494
<212> DNA
<213> 蒺藜苜蓿(Medicago truncatula)
<220>
<221> misc_feature
<223> 3GT3
<400> 40
atggaaggtg ttgaagttga acaaccattg aaagtttact tcattccatt tcttgcatct 60
ggacatatga tccctctttt tgacatagca actatgtttg catcccgtgg ccagcaagta 120
acagtcatca ccactcccgc caacgcaaaa tcccttacca aatctctctc atccgacgct 180
ccttcattcc ttcgtcttca caccgttgac ttcccctccc aacaagtcgg cctccctgaa 240
ggtattgaat ctatgtcttc aactacggac cccaccacca cttggaagat ccatactggc 300
gcgatgctcc ttaaagaacc tattggggat ttcattgaga atgatccacc ggattgtatc 360
atctccgact ccacgtaccc atgggttaat gacttggccg ataagtttca gatcccaaac 420
atcacattca atggattgtg cctttttgct gtctccctcg tggaaaccct caaaacaaac 480
aatttactta agtctcagac agattctgat tcggattcaa gttcctttgt tgttccaaat 540
tttcctcacc atatcacctt gtgtggaaaa ccgccaaagg taatcggtat attcatggga 600
atgatgcttg agacggtgct taaaagtaaa gcactaatca tcaacaactt cagtgaactt 660
gatggagaag agtgcataca acactacgag aaagccacgg gtcacaaggt ttggcatctt 720
ggtccaactt ctcttattcg caaaactgct caagagaaat cagagagggg aaatgagggt 780
gctgtgaatg tgcacgagag cctgagttgg ctcgattcag agagagttaa ctcagtgttg 840
tacatatgtt ttggaagcat caactatttt tctgataaac aactatacga gatggcatgt 900
gcgatagaag catccggtca cccattcata tgggttgttc ctgagaagaa agggaaagaa 960
gatgagagcg aagaagagaa agaaaagtgg ttaccgaagg gatttgaaga gagaaatatc 1020
gggaagaagg gtttgatcat taggggttgg gccccacagg ttaagatatt gagccaccct 1080
gcagtggggg gatttatgac gcattgcggg gggaactcaa ccgtagaggc tgttagtgcg 1140
ggagttccaa tgataacgtg gccggttcat ggagatcaat tctacaatga gaaactgata 1200
acacaattcc gaggaattgg agttgaagtc ggtgcaacag aatggtgtac gagtggtgtc 1260
gcggagagaa agaagttagt gagcagagat agcatagaga aggctgtaag gagattgatg 1320
gacggtggtg atgaagctga aaatatcagg ctacgtgctc gagagtttgg agaaaaagct 1380
atacaagcta ttcaagaagg tggctcgtct tataataatt tgttggcttt gattgacgaa 1440
cttaaaagat cgagagacct taaaagattg agagacctca agctggatga ttaa 1494
<210> 41
<211> 497
<212> PRT
<213> 蒺藜苜蓿(Medicago truncatula)
<220>
<221> misc_feature
<223> 3GT3
<400> 41
Met Glu Gly Val Glu Val Glu Gln Pro Leu Lys Val Tyr Phe Ile Pro
1 5 10 15
Phe Leu Ala Ser Gly His Met Ile Pro Leu Phe Asp Ile Ala Thr Met
20 25 30
Phe Ala Ser Arg Gly Gln Gln Val Thr Val Ile Thr Thr Pro Ala Asn
35 40 45
Ala Lys Ser Leu Thr Lys Ser Leu Ser Ser Asp Ala Pro Ser Phe Leu
50 55 60
Arg Leu His Thr Val Asp Phe Pro Ser Gln Gln Val Gly Leu Pro Glu
65 70 75 80
Gly Ile Glu Ser Met Ser Ser Thr Thr Asp Pro Thr Thr Thr Trp Lys
85 90 95
Ile His Thr Gly Ala Met Leu Leu Lys Glu Pro Ile Gly Asp Phe Ile
100 105 110
Glu Asn Asp Pro Pro Asp Cys Ile Ile Ser Asp Ser Thr Tyr Pro Trp
115 120 125
Val Asn Asp Leu Ala Asp Lys Phe Gln Ile Pro Asn Ile Thr Phe Asn
130 135 140
Gly Leu Cys Leu Phe Ala Val Ser Leu Val Glu Thr Leu Lys Thr Asn
145 150 155 160
Asn Leu Leu Lys Ser Gln Thr Asp Ser Asp Ser Asp Ser Ser Ser Phe
165 170 175
Val Val Pro Asn Phe Pro His His Ile Thr Leu Cys Gly Lys Pro Pro
180 185 190
Lys Val Ile Gly Ile Phe Met Gly Met Met Leu Glu Thr Val Leu Lys
195 200 205
Ser Lys Ala Leu Ile Ile Asn Asn Phe Ser Glu Leu Asp Gly Glu Glu
210 215 220
Cys Ile Gln His Tyr Glu Lys Ala Thr Gly His Lys Val Trp His Leu
225 230 235 240
Gly Pro Thr Ser Leu Ile Arg Lys Thr Ala Gln Glu Lys Ser Glu Arg
245 250 255
Gly Asn Glu Gly Ala Val Asn Val His Glu Ser Leu Ser Trp Leu Asp
260 265 270
Ser Glu Arg Val Asn Ser Val Leu Tyr Ile Cys Phe Gly Ser Ile Asn
275 280 285
Tyr Phe Ser Asp Lys Gln Leu Tyr Glu Met Ala Cys Ala Ile Glu Ala
290 295 300
Ser Gly His Pro Phe Ile Trp Val Val Pro Glu Lys Lys Gly Lys Glu
305 310 315 320
Asp Glu Ser Glu Glu Glu Lys Glu Lys Trp Leu Pro Lys Gly Phe Glu
325 330 335
Glu Arg Asn Ile Gly Lys Lys Gly Leu Ile Ile Arg Gly Trp Ala Pro
340 345 350
Gln Val Lys Ile Leu Ser His Pro Ala Val Gly Gly Phe Met Thr His
355 360 365
Cys Gly Gly Asn Ser Thr Val Glu Ala Val Ser Ala Gly Val Pro Met
370 375 380
Ile Thr Trp Pro Val His Gly Asp Gln Phe Tyr Asn Glu Lys Leu Ile
385 390 395 400
Thr Gln Phe Arg Gly Ile Gly Val Glu Val Gly Ala Thr Glu Trp Cys
405 410 415
Thr Ser Gly Val Ala Glu Arg Lys Lys Leu Val Ser Arg Asp Ser Ile
420 425 430
Glu Lys Ala Val Arg Arg Leu Met Asp Gly Gly Asp Glu Ala Glu Asn
435 440 445
Ile Arg Leu Arg Ala Arg Glu Phe Gly Glu Lys Ala Ile Gln Ala Ile
450 455 460
Gln Glu Gly Gly Ser Ser Tyr Asn Asn Leu Leu Ala Leu Ile Asp Glu
465 470 475 480
Leu Lys Arg Ser Arg Asp Leu Lys Arg Leu Arg Asp Leu Lys Leu Asp
485 490 495
Asp
<210> 42
<211> 1374
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> 3GT4
<400> 42
atggagagag aaatgttgag caaaactcac attatgttca tcccattccc agctcaaggc 60
cacatgagcc caatgatgca attcgccaag cgtttagcct ggaaaggcct gcgaatcacg 120
atagttcttc cggctcaaat tcgagatttc atgcaaataa ccaacccatt gatcaacact 180
gagtgcatct cctttgattt tgataaagac gatgggatgc catacagcat gcaggcttat 240
atgggagttg taaaactcaa ggtcacaaat aaactgagtg acctactcga gaagcaaaga 300
acaaatggct accctgttaa tttgctagtg gttgattcat tatatccatc tcgggtagaa 360
atgtgccacc aacttggggt aaaaggagct ccatttttca ctcactcttg tgctgttggt 420
gccatttatt ataatgctcg cttagggaaa ttgaagatac ctcctgagga agggttgact 480
tctgtttcat tgccttcaat tccattgttg gggagagatg atttgccaat tattaggact 540
ggcacctttc ctgatctctt tgagcatttg gggaatcagt tttcagatct tgataaagcg 600
gattggatct ttttcaatac ttttgataag cttgaaaatg aggaagcaaa atggctatct 660
agccaatggc caattacatc catcggacca ttaatccctt caatgtactt agacaaacaa 720
ttaccaaatg acaaagacaa tggcattaat ttctacaagg cagacgtcgg atcgtgcatc 780
aagtggctag acgccaaaga ccctggctcg gtagtctacg cctcattcgg gagcgtgaag 840
cacaacctcg gcgatgacta catggacgaa gtagcatggg gcttgttaca tagcaaatat 900
cacttcatat gggttgttat agaatccgaa cgtacaaagc tctctagcga tttcttggca 960
gaggcagagg cagaggaaaa aggcctaata gtgagttggt gccctcaact ccaagttttg 1020
tcacataaat ctatagggag ttttatgact cattgtggtt ggaactcgac ggttgaggca 1080
ttgagtttgg gcgtgccaat ggtggcactg ccacaacagt ttgatcagcc tgctaatgcc 1140
aagtatatcg tggatgtatg gcaaattggg gttcgggttc cgattggtga agagggggtt 1200
gttttgaggg gagaagttgc taactgtata aaggatgtta tggaggggga aataggggat 1260
gagcttagag ggaatgcttt gaaatggaag gggttggctg tggaggcaat ggagaaaggg 1320
ggtagctctg ataagaatat tgatgagttc atttcaaagc ttgtttcctc ctga 1374
<210> 43
<211> 457
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> 3GT4
<400> 43
Met Glu Arg Glu Met Leu Ser Lys Thr His Ile Met Phe Ile Pro Phe
1 5 10 15
Pro Ala Gln Gly His Met Ser Pro Met Met Gln Phe Ala Lys Arg Leu
20 25 30
Ala Trp Lys Gly Leu Arg Ile Thr Ile Val Leu Pro Ala Gln Ile Arg
35 40 45
Asp Phe Met Gln Ile Thr Asn Pro Leu Ile Asn Thr Glu Cys Ile Ser
50 55 60
Phe Asp Phe Asp Lys Asp Asp Gly Met Pro Tyr Ser Met Gln Ala Tyr
65 70 75 80
Met Gly Val Val Lys Leu Lys Val Thr Asn Lys Leu Ser Asp Leu Leu
85 90 95
Glu Lys Gln Arg Thr Asn Gly Tyr Pro Val Asn Leu Leu Val Val Asp
100 105 110
Ser Leu Tyr Pro Ser Arg Val Glu Met Cys His Gln Leu Gly Val Lys
115 120 125
Gly Ala Pro Phe Phe Thr His Ser Cys Ala Val Gly Ala Ile Tyr Tyr
130 135 140
Asn Ala Arg Leu Gly Lys Leu Lys Ile Pro Pro Glu Glu Gly Leu Thr
145 150 155 160
Ser Val Ser Leu Pro Ser Ile Pro Leu Leu Gly Arg Asp Asp Leu Pro
165 170 175
Ile Ile Arg Thr Gly Thr Phe Pro Asp Leu Phe Glu His Leu Gly Asn
180 185 190
Gln Phe Ser Asp Leu Asp Lys Ala Asp Trp Ile Phe Phe Asn Thr Phe
195 200 205
Asp Lys Leu Glu Asn Glu Glu Ala Lys Trp Leu Ser Ser Gln Trp Pro
210 215 220
Ile Thr Ser Ile Gly Pro Leu Ile Pro Ser Met Tyr Leu Asp Lys Gln
225 230 235 240
Leu Pro Asn Asp Lys Asp Asn Gly Ile Asn Phe Tyr Lys Ala Asp Val
245 250 255
Gly Ser Cys Ile Lys Trp Leu Asp Ala Lys Asp Pro Gly Ser Val Val
260 265 270
Tyr Ala Ser Phe Gly Ser Val Lys His Asn Leu Gly Asp Asp Tyr Met
275 280 285
Asp Glu Val Ala Trp Gly Leu Leu His Ser Lys Tyr His Phe Ile Trp
290 295 300
Val Val Ile Glu Ser Glu Arg Thr Lys Leu Ser Ser Asp Phe Leu Ala
305 310 315 320
Glu Ala Glu Ala Glu Glu Lys Gly Leu Ile Val Ser Trp Cys Pro Gln
325 330 335
Leu Gln Val Leu Ser His Lys Ser Ile Gly Ser Phe Met Thr His Cys
340 345 350
Gly Trp Asn Ser Thr Val Glu Ala Leu Ser Leu Gly Val Pro Met Val
355 360 365
Ala Leu Pro Gln Gln Phe Asp Gln Pro Ala Asn Ala Lys Tyr Ile Val
370 375 380
Asp Val Trp Gln Ile Gly Val Arg Val Pro Ile Gly Glu Glu Gly Val
385 390 395 400
Val Leu Arg Gly Glu Val Ala Asn Cys Ile Lys Asp Val Met Glu Gly
405 410 415
Glu Ile Gly Asp Glu Leu Arg Gly Asn Ala Leu Lys Trp Lys Gly Leu
420 425 430
Ala Val Glu Ala Met Glu Lys Gly Gly Ser Ser Asp Lys Asn Ile Asp
435 440 445
Glu Phe Ile Ser Lys Leu Val Ser Ser
450 455
<210> 44
<211> 24
<212> DNA
<213> oligonucleotides
<400> 44
atggaaggtg ttgaagttga acaa 24
<210> 45
<211> 24
<212> DNA
<213> oligonucleotides
<400> 45
ttaatcatcc agcttgaggt ctct 24
<210> 46
<211> 25
<212> DNA
<213> oligonucleotides
<400> 46
tacataaata tataatacat aggca 25
<210> 47
<211> 25
<212> DNA
<213> oligonucleotides
<400> 47
aaaatacata caaaatcttg aaata 25
<210> 48
<211> 45
<212> DNA
<213> oligonucleotides
<400> 48
actttaagaa ggagatatac catggaaggt gttgaagttg aacaa 45
<210> 49
<211> 45
<212> DNA
<213> 人工序列
<400> 49
ctcgagtgcg gccgcaagct tatcatccag cttgaggtct ctcaa 45
<210> 50
<211> 46
<212> DNA
<213> oligonucleotides
<400> 50
actttaagaa ggagatatac catggagaga gaaatgttga gcaaaa 46
<210> 51
<211> 46
<212> DNA
<213> oligonucleotides
<400> 51
ctcgagtgcg gccgcaagct tggaggaaac aagctttgaa atgaac 46
<210> 52
<211> 27
<212> DNA
<213> oligonucleotides
<400> 52
ggtatatctc cttcttaaag ttaaaca 27
<210> 53
<211> 27
<212> DNA
<213> oligonucleotides
<400> 53
aagcttgcgg ccgcactcga gcaccac 27
<210> 54
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-4
<400> 54
atggataacc aaaaaggtag aatcagtata gcgttgctac catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattgggg gagcagcctc cttttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agttattact tgattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatattttg atttttattc tactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagtttat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagaggggtt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattacgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
catcttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagggattg cagaggtaat aagaaaagtc 1200
gttgtggaga aaagtgggga ggttatgagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagaaag gagaggaaga gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 55
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-4
<400> 55
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Phe Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Met Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Glu Glu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 56
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-5
<400> 56
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattgggg gagcagcctc cttttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agttattact tgattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatatatcg atttgctttc cactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagattat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagagggatt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattgcgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
catcttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagggattg cagaggtaat aagaaaagtc 1200
gttgtggaga aaagtgggga ggttatcagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagatag gagagcaatt gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 57
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-5
<400> 57
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Phe Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 58
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-6
<400> 58
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttatttc 420
ctaaccacgg cagcagccag ctcttccatt ggcctacatg ctttcaaaaa cccaggtgaa 480
aaatacccat ttccagattt ttatgataac agtaataata cccctgaacc accttctgca 540
gataacatga agctacttca tgattttatc gcttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttataga actagaaggg aaatatatcg atttgctttc cactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagattat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagagggatt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattgcgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
catcttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagatattg caggggtaat aagaaaagtc 1200
gtggtggaga aaagtgggga ggttatcagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagatag gagagcaatt gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 59
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-6
<400> 59
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala
130 135 140
Ala Ala Ser Ser Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Pro Asp Phe Tyr Asp Asn Ser Asn Asn Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Ile Glu Leu
195 200 205
Glu Gly Lys Tyr Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 60
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-7
<400> 60
atggataacc aaaaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcaaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagag tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atgatcccac tcagaaacgc ctttgataca 300
gcaggcccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattggtg gagcagcctc ctcttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agctattaat taattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatattttg attttttttc cactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagtttat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagtg ttttctctcc aatgaggaat tggaagaagt agcgattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagaggggtt tgttcaaagg gtaggagaca gaggattggt tgtggaggag 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggaatt ctattacgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
cattttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagggattg cagaggtaat aagaaaagtc 1200
gttgtggaga aaagtgggga ggttatcagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagaaag gagagcaaga gattgatagg gtagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 61
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<220>
<221> misc_feature
<223> gGT29-7
<400> 61
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Val Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Ile Pro Leu Arg Asn
85 90 95
Ala Phe Asp Thr Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Ser Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Ile Asn Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Phe Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Cys Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Glu
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Asn Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Val Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 62
<211> 26
<212> DNA
<213> 人工序列
<400> 62
acagcaagag agagacacag agttca 26
<210> 63
<211> 23
<212> DNA
<213> oligonucleotides
<400> 63
gttcaaagcc caacctaagc gca 23
<210> 64
<211> 25
<212> DNA
<213> oligonucleotides
<400> 64
atgaaattat acagagaggg agaga 25
<210> 65
<211> 25
<212> DNA
<213> oligonucleotides
<400> 65
tggttttctt cacaacacaa agtac 25
<210> 66
<211> 43
<212> DNA
<213> oligonucleotides
<400> 66
ctggtgccgc gcggcagcat ggataaccaa aagggtagaa tca 43
<210> 67
<211> 43
<212> DNA
<213> oligonucleotides
<400> 67
ctggtgccgc gcggcagcat ggataaccaa aaaggtagaa tca 43
<210> 68
<211> 43
<212> DNA
<213> oligonucleotides
<400> 68
tgcggccgca agcttgtctt gttcatcttt cttcttctta caa 43
<210> 69
<211> 30
<212> DNA
<213> oligonucleotides
<400> 69
gctgccgcgc ggcaccaggc cgctgctgtg 30
<210> 70
<211> 30
<212> DNA
<213> oligonucleotides
<400> 70
tccgtcgaca agcttgcggc cgcactcgag 30
<210> 71
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<400> 71
atggataacc aagaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcaaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aagaataagg attcctctgc ttctataaaa 180
ctagtagagc ttcatcttcc atcttcccct gatcttcctc ctcactacca caccacaaat 240
ggcctccctt cccatctcat ggtcccactc agaaacgcct ttgaaacagc agcccccacc 300
ttctctgaaa tccttaaaac cttaaaccct gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttgtttcat aattggggga 420
gcagcctcct tttccatgag cctacatagt ttcaaaaacc caggtgaaaa atacccattt 480
ctagattttg atgataacag taatattacc cctgaaccac cttcagcaga taacatgaag 540
ttattacttg attttatgac ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttagagaac tagaagggaa atattttgat ttttattcta ctttatctga taaaactttg 660
gttcctgttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagtttataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
atggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg ggttttacca 900
gaggggtttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attacggaga gtatgaagtt tggggttcca gtatttgcca tggccaggca ttttgatcag 1080
cctttgaatg ctaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgaa 1140
aatgggaagt ataagagaga agatattgca ggggtaataa gaaaagtcgt ggtggagaaa 1200
agtggggagg ttatcaggag gaaagcaagg gaattgagtg agaaaatgaa agagatagga 1260
gagcaattga ttgatagggc agtggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gaacaatag 1329
<210> 72
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<400> 72
Met Asp Asn Gln Glu Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn Ala Phe Glu Thr
85 90 95
Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly Ala Ala Ser Phe
130 135 140
Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu Glu Gly Lys Tyr
195 200 205
Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Val Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro Glu Gly Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Thr Glu Ser Met Lys Phe Gly Val Pro Val Phe
340 345 350
Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Ala Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440
<210> 73
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<400> 73
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattgggg gagcagcctc cttttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agttattact tgattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatattttg atttttattc tactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagtttat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagagggatt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattgcgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
catcttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagatattg caggggtaat aagaaaagtc 1200
gtggtggaga aaagtgggga ggttatcagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagatag gagagcaatt gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 74
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<400> 74
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Phe Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 75
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<400> 75
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactat 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcataaacgc ctttgaaaca 300
gcaggcccca ccttctctga aatccttaaa accttaaacc ccgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttatttc 420
ctaaccacgg cagcagccag ctcttccatt ggcctacatg ctttcaaaaa cccaggtgaa 480
aaatacccat ttccagattt ttatgataac agtaataata cccctgaacc accttctgca 540
gataacatga agctacttca tgattttatc gcttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttataga actagaaggg aaatatatcg atttgctttc cactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagattat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagagggatt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattgcgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
catcttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagggattg cagaggtaat aagaaaagtc 1200
gttgtggaga aaagtgggga ggttatgagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagaaag gagaggaaga gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgcacaata g 1341
<210> 76
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<400> 76
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Ile Asn
85 90 95
Ala Phe Glu Thr Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala
130 135 140
Ala Ala Ser Ser Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Pro Asp Phe Tyr Asp Asn Ser Asn Asn Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Ile Glu Leu
195 200 205
Glu Gly Lys Tyr Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Met Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Glu Glu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Ala Gln
435 440 445
<210> 77
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<400> 77
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattgggg gagcagcctc cttttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agttattact tgattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatattttg atttttattc tactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagtttat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagaggggtt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattacgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
cattttgatc agcctttgaa tgctaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagatattg caggggtaat aagaaaagtc 1200
gtggtggaga aaagtgggga ggttatcagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagatag gagagcaatt gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 78
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<400> 78
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Phe Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Ala
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 79
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<400> 79
atggataacc aagaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcaaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aagaataagg attcctctgc ttctataaaa 180
ctagttgagc ttcatcttcc atcttcccct gatcttcctc ctcactatca caccacaaat 240
ggcctccctt cccatctcat ggtcccactc ataaacgcct ttgaaacagc aggccccacc 300
ttctctgaaa tccttaaaac cttaaacccc gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttatttcct aaccacggca 420
gcagccagct cttccattgg cctacatgct ttcaaaaacc caggtgaaaa atacccattt 480
ccagattttt atgataacag taataatacc cctgaaccac cttctgcaga taacatgaag 540
ctacttcatg attttatcgc ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttatagaac tagaagggaa atatatcgat ttgctttcca ctttatctga taaaactttg 660
gttccttttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagattataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
atggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg ggttttacca 900
gagggatttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attgcggaga gtatgaagtt tggggttcca gtaattgcca tggccaggca tcttgatcag 1080
cctttgaatg gtaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgaa 1140
aatgggaagt ataagagaga agggattgca gaggtaataa gaaaagtcgt tgtggagaaa 1200
agtggggagg ttatgaggag gaaagcaagg gaattgagtg agaaaatgaa agagaaagga 1260
gaggaagaga ttgatagggc agtggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gcacaatag 1329
<210> 80
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<400> 80
Met Asp Asn Gln Glu Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Val Pro Leu Ile Asn Ala Phe Glu Thr
85 90 95
Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala Ala Ala Ser Ser
130 135 140
Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Pro Asp Phe Tyr Asp Asn Ser Asn Asn Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Ile Glu Leu Glu Gly Lys Tyr
195 200 205
Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Phe Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro Glu Gly Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly Val Pro Val Ile
340 345 350
Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Met Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Lys Gly Glu Glu Glu Ile Asp Arg Ala Val Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Ala Gln
435 440
<210> 81
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<400> 81
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattgggg gagcagcctc cttttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agttattact tgattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatattttg atttttattc tactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagtttat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagaggggtt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattacgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
cattttgatc agcctttgaa tgctaagctg gcggcggagg ttggtgtggt catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagatattg caggggtaat aagaaaagtc 1200
gtggtggaga aaagtgggga ggttatcagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagatag gagagcaatt gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgaacaata g 1341
<210> 82
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<400> 82
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Phe Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Ala
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Val Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 83
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<400> 83
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcaggcccca ccttctctga aatccttaaa accttaaacc ccgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttatttc 420
ctaaccacgg cagcagccag ctcttccatt ggcctacatg ctttcaaaaa cccaggtgaa 480
aaatacccat ttccagattt ttatgataac agtaataata cccctgaacc accttctgca 540
gataacatga agctacttca tgattttatc gcttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttataga actagaaggg aaatatatcg atttgctttc cactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagattat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagagggatt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattgcgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
catcttgatc agcctttgaa tggtaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagggattg cagaggtaat aagaaaagtc 1200
gttgtggaga aaagtgggga ggttatgagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagaaag gagaggaaga gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgcacaata g 1341
<210> 84
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<400> 84
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala
130 135 140
Ala Ala Ser Ser Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Pro Asp Phe Tyr Asp Asn Ser Asn Asn Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Ile Glu Leu
195 200 205
Glu Gly Lys Tyr Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Met Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Glu Glu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Ala Gln
435 440 445
<210> 85
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<400> 85
atggataacc aaaaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcgaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aaggataagg attcctctgc ttctataaaa 180
ctagttgagc ttcatcttcc atcttcccct gatcttcctc ctcactatca caccacaaat 240
ggcctccctt cccatctcat gctcccactc agaaacgcct ttgaaactgc aggccccacc 300
ttctctgaaa tccttaaaac cttaaacccc gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttatttcct aaccacggca 420
gcagccagct cttccattgg cctacatgct ttcaaaaacc caggtgaaaa atacccattt 480
ccagattttt atgataacag taatattacc cctgaaccac cttctgcaga taacatgaag 540
ctacttcatg attttatcgc ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttagagaac tagaagggaa atatattgat ttgctttcca ctttatctga taaaactttg 660
gttcctgttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagattataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
acggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg gattttacca 900
gagcggtttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attgcggaga gtatgaagtt tggggttcca gtaattgcca tggccaggca tcttgatcag 1080
cctttgaatg gtaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgag 1140
aatgggaagt ataagagaga agggattgca gaggtaataa gaaaagtggt tgtggagaaa 1200
agtggggagg ttatcaggag gaaagcaagg gagttgagtg agaaaatgaa agagaaagga 1260
gagcaagaga ttgatagggc attggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gaacaatag 1329
<210> 86
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<400> 86
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asp Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Leu Pro Leu Arg Asn Ala Phe Glu Thr
85 90 95
Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala Ala Ala Ser Ser
130 135 140
Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Pro Asp Phe Tyr Asp Asn Ser Asn Ile Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu Glu Gly Lys Tyr
195 200 205
Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Val Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Thr Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Ile Leu Pro Glu Arg Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly Val Pro Val Ile
340 345 350
Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Ala Leu Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440
<210> 87
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<400> 87
atggataacc aaaaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcgaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aaggataagg attcctctgc ttctataaaa 180
ctagttgagc ttcatcttcc atcttcccct gatcttcctc ctcactatca caccacaaat 240
ggcctccctt cccatctcat gctcccactc agaaacgcct ttgaaactgc aggccccacc 300
ttctctgaaa tccttaaaac cttaaacccc gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttatttcct aaccacggca 420
gcagccagct cttccattgg cctacatgct ttcaaaaacc caggtgaaaa atacccattt 480
ccagattttt atgataacag taatattacc cctgaaccac cttctgcaga taacatgaag 540
ctacttcatg attttatcgc ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttagagaac tagaagggaa atatattgat ttgctttcca ctttatctga taaaactttg 660
gttcctgttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagattataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
acggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg gattttacca 900
gaggggtttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attgcggaga gtatgaagtt tggggttcca gtaattgcca tggccaggca tcttgatcag 1080
cctttgaatg gtaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgag 1140
aatgggaagt ataagagaga agggattgca gaggtaataa gaaaagtggt tgtggagaaa 1200
agtggggagg ttatcaggag gaaagcaagg gagttgagtg agaaaatgaa agagaaagga 1260
gagcaagaga ttgatagggt agtggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gaacaatag 1329
<210> 88
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<400> 88
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asp Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Leu Pro Leu Arg Asn Ala Phe Glu Thr
85 90 95
Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Tyr Phe Leu Thr Thr Ala Ala Ala Ser Ser
130 135 140
Ser Ile Gly Leu His Ala Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Pro Asp Phe Tyr Asp Asn Ser Asn Ile Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu His Asp Phe Ile Ala Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu Glu Gly Lys Tyr
195 200 205
Ile Asp Leu Leu Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Val Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Ile Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Thr Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Ile Leu Pro Glu Gly Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Ala Glu Ser Met Lys Phe Gly Val Pro Val Ile
340 345 350
Ala Met Ala Arg His Leu Asp Gln Pro Leu Asn Gly Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Val Val Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440
<210> 89
<211> 1329
<212> DNA
<213> 人参(Panax ginseng)
<400> 89
atggataacc aagaaggtag aatcagtata gcgttgctac catttttagc ccatggtcac 60
atatctccct tctttgagct agccaaacaa ctcgcaaaaa gaaattgcaa tgttttcctc 120
tgttctaccc caatcaatct tagctccatc aagaataagg attcctctgc ttctataaaa 180
ctagttgagc ttcatcttcc atcttcccct gatcttcctc ctcactatca caccacaaat 240
ggcctccctt cccatctcat ggtcccactc ataaacgcct ttgaaacagc aggccccacc 300
ttctctgaaa tccttaaaac cttaaaccct gatttgctta tttatgattt caatccctca 360
tgggcaccgg agatcgcttc gtctcacaat attccggcag tttgtttcat aattggggga 420
gcagcctcct tttccatgag cctacatagt ttcaaaaacc caggtgaaaa atacccattt 480
ctagattttg atgataacag taatattacc cctgaaccac cttcagcaga taacatgaag 540
ttattacttg attttatgac ttgtttcgaa cgatcttgcg acattatttt gattaagagt 600
tttagagaac tagaagggaa atattttgat ttttattcta ctttatctga taaaactttg 660
gttcctgttg gtccactcgt tcaagatcct atgggccata atgaagatcc aaaaacagag 720
cagtttataa actggcttga caaaagggct gaatctacag tggtgtttgt ctgctttgga 780
agtgagtatt ttctctccaa tgaggaattg gaagaagtag caattgggct agagattagc 840
atggttaatt tcatatgggc tgtgagatta attgaaggag agaaaaaagg ggttttacca 900
gaggggtttg ttcaaagggt aggagacaga ggattggttg tggaggggtg ggctccacag 960
gcaagaattt taggacattc aagcaccggt gggtttgtga gccattgtgg gtggagttct 1020
attacggaga gtatgaagtt tggggttcca gtaattgcca tggccaggca ttttgatcag 1080
cctttgaatg ctaagctggc ggcggaggtt ggtgtgggca tggaggttgt gagagatgaa 1140
aatgggaagt ataagagaga agatattgca ggggtaataa gaaaagtcgt ggtggagaaa 1200
agtggggagg ttatcaggag gaaagcaagg gaattgagtg agaaaatgaa agagatagga 1260
gagcaattga ttgatagggc agtggaggag ctagtacaaa tttgtaagaa gaagaaagat 1320
gaacaatag 1329
<210> 90
<211> 442
<212> PRT
<213> 人参(Panax ginseng)
<400> 90
Met Asp Asn Gln Glu Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Lys Asp Ser Ser Ala Ser Ile Lys Leu Val Glu Leu
50 55 60
His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr His Thr Thr Asn
65 70 75 80
Gly Leu Pro Ser His Leu Met Val Pro Leu Ile Asn Ala Phe Glu Thr
85 90 95
Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu Asn Pro Asp Leu
100 105 110
Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu Ile Ala Ser Ser
115 120 125
His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly Ala Ala Ser Phe
130 135 140
Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu Lys Tyr Pro Phe
145 150 155 160
Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu Pro Pro Ser Ala
165 170 175
Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys Phe Glu Arg Ser
180 185 190
Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu Glu Gly Lys Tyr
195 200 205
Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu Val Pro Val Gly
210 215 220
Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp Pro Lys Thr Glu
225 230 235 240
Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser Thr Val Val Phe
245 250 255
Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu Glu Leu Glu Glu
260 265 270
Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe Ile Trp Ala Val
275 280 285
Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro Glu Gly Phe Val
290 295 300
Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly Trp Ala Pro Gln
305 310 315 320
Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe Val Ser His Cys
325 330 335
Gly Trp Ser Ser Ile Thr Glu Ser Met Lys Phe Gly Val Pro Val Ile
340 345 350
Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Ala Lys Leu Ala Ala
355 360 365
Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu Asn Gly Lys Tyr
370 375 380
Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val Val Val Glu Lys
385 390 395 400
Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu Ser Glu Lys Met
405 410 415
Lys Glu Ile Gly Glu Gln Leu Ile Asp Arg Ala Val Glu Glu Leu Val
420 425 430
Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440
<210> 91
<211> 1341
<212> DNA
<213> 人参(Panax ginseng)
<400> 91
atggataacc aaaagggtag aatcagtata gttatgctgc catttttagc ccatggccac 60
atttctccat tctttgagct agccaagcat ctctcaaaaa gaaattgtaa tatattcctc 120
tgttctaccc caatcaatct tagctccatc aagaacagaa tatctgataa ggattcctct 180
gcttctataa aactagtaga gcttcatctt ccatcttccc ctgatcttcc tcctcactac 240
cacaccacaa atggcctccc ttcccatctc atggtcccac tcagaaacgc ctttgaaaca 300
gcagccccca ccttctctga aatccttaaa accttaaacc ctgatttgct tatttatgat 360
ttcaatccct catgggcacc ggagatcgct tcgtctcaca atattccggc agtttgtttc 420
ataattgggg gagcagcctc cttttccatg agcctacata gtttcaaaaa cccaggtgaa 480
aaatacccat ttctagattt tgatgataac agtaatatta cccctgaacc accttcagca 540
gataacatga agttattact tgattttatg acttgtttcg aacgatcttg cgacattatt 600
ttgattaaga gttttagaga actagaaggg aaatattttg atttttattc tactttatct 660
gataaaactt tggttcctgt tggtccactc gttcaagatc ctatgggcca taatgaagat 720
ccaaaaacag agcagtttat aaactggctt gacaaaaggg ctgaatctac agtggtgttt 780
gtctgctttg gaagtgagta ttttctctcc aatgaggaat tggaagaagt agcaattggg 840
ctagagatta gcatggttaa tttcatatgg gctgtgagat taattgaagg agagaaaaaa 900
ggggttttac cagaggggtt tgttcaaagg gtaggagaca gaggattggt tgtggagggg 960
tgggctccac aggcaagaat tttaggacat tcaagcaccg gtgggtttgt gagccattgt 1020
gggtggagtt ctattacgga gagtatgaag tttggggttc cagtaattgc catggccagg 1080
cattttgatc agcctttgaa tgctaagctg gcggcggagg ttggtgtggg catggaggtt 1140
gtgagagatg aaaatgggaa gtataagaga gaagatattg caggggtaat aagaaaagtc 1200
gtggtggaga aaagtgggga ggttatgagg aggaaagcaa gggaattgag tgagaaaatg 1260
aaagagaaag gagaggaaga gattgatagg gcagtggagg agctagtaca aatttgtaag 1320
aagaagaaag atgcacaata g 1341
<210> 92
<211> 446
<212> PRT
<213> 人参(Panax ginseng)
<400> 92
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Val Met Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys His Leu Ser
20 25 30
Lys Arg Asn Cys Asn Ile Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Ile Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Val Pro Leu Arg Asn
85 90 95
Ala Phe Glu Thr Ala Ala Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Phe Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Leu Asp Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Tyr Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Tyr Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Gly
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Ser Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Ala
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Asp Ile Ala Gly Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Met Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Glu Glu Ile Asp Arg Ala Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Ala Gln
435 440 445
<210> 93
<211> 446
<212> PRT
<213> 突变蛋白
<400> 93
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Val Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Ile Pro Leu Arg Asn
85 90 95
Ala Phe Asp Thr Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Ser Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Ile Asn Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Phe Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Cys Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Glu
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Gly Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Ala Arg His Phe Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Val Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 94
<211> 446
<212> PRT
<213> 突变蛋白
<400> 94
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Val Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Ile Pro Leu Arg Asn
85 90 95
Ala Phe Asp Thr Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Ser Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Ile Asn Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Phe Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Cys Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Glu
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Asn Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Pro Arg His Phe Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Val Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445
<210> 95
<211> 446
<212> PRT
<213> 突变蛋白
<400> 95
Met Asp Asn Gln Lys Gly Arg Ile Ser Ile Ala Leu Leu Pro Phe Leu
1 5 10 15
Ala His Gly His Ile Ser Pro Phe Phe Glu Leu Ala Lys Gln Leu Ala
20 25 30
Lys Arg Asn Cys Asn Val Phe Leu Cys Ser Thr Pro Ile Asn Leu Ser
35 40 45
Ser Ile Lys Asn Arg Val Ser Asp Lys Asp Ser Ser Ala Ser Ile Lys
50 55 60
Leu Val Glu Leu His Leu Pro Ser Ser Pro Asp Leu Pro Pro His Tyr
65 70 75 80
His Thr Thr Asn Gly Leu Pro Ser His Leu Met Ile Pro Leu Arg Asn
85 90 95
Ala Phe Asp Thr Ala Gly Pro Thr Phe Ser Glu Ile Leu Lys Thr Leu
100 105 110
Asn Pro Asp Leu Leu Ile Tyr Asp Phe Asn Pro Ser Trp Ala Pro Glu
115 120 125
Ile Ala Ser Ser His Asn Ile Pro Ala Val Cys Phe Ile Ile Gly Gly
130 135 140
Ala Ala Ser Ser Ser Met Ser Leu His Ser Phe Lys Asn Pro Gly Glu
145 150 155 160
Lys Tyr Pro Phe Leu Asp Phe Asp Asp Asn Ser Asn Ile Thr Pro Glu
165 170 175
Pro Pro Ser Ala Asp Asn Met Lys Leu Leu Ile Asn Phe Met Thr Cys
180 185 190
Phe Glu Arg Ser Cys Asp Ile Ile Leu Ile Lys Ser Phe Arg Glu Leu
195 200 205
Glu Gly Lys Tyr Phe Asp Phe Phe Ser Thr Leu Ser Asp Lys Thr Leu
210 215 220
Val Pro Val Gly Pro Leu Val Gln Asp Pro Met Gly His Asn Glu Asp
225 230 235 240
Pro Lys Thr Glu Gln Phe Ile Asn Trp Leu Asp Lys Arg Ala Glu Ser
245 250 255
Thr Val Val Phe Val Cys Phe Gly Ser Glu Cys Phe Leu Ser Asn Glu
260 265 270
Glu Leu Glu Glu Val Ala Ile Gly Leu Glu Ile Ser Met Val Asn Phe
275 280 285
Ile Trp Ala Val Arg Leu Ile Glu Gly Glu Lys Lys Gly Val Leu Pro
290 295 300
Glu Gly Phe Val Gln Arg Val Gly Asp Arg Gly Leu Val Val Glu Glu
305 310 315 320
Trp Ala Pro Gln Ala Arg Ile Leu Gly His Ser Ser Thr Gly Gly Phe
325 330 335
Val Ser His Cys Gly Trp Gly Ser Ile Thr Glu Ser Met Lys Phe Gly
340 345 350
Val Pro Val Ile Ala Met Pro Arg His Phe Asp Gln Pro Leu Asn Gly
355 360 365
Lys Leu Ala Ala Glu Val Gly Val Gly Met Glu Val Val Arg Asp Glu
370 375 380
Asn Gly Lys Tyr Lys Arg Glu Gly Ile Ala Glu Val Ile Arg Lys Val
385 390 395 400
Val Val Glu Lys Ser Gly Glu Val Ile Arg Arg Lys Ala Arg Glu Leu
405 410 415
Ser Glu Lys Met Lys Glu Lys Gly Glu Gln Glu Ile Asp Arg Val Val
420 425 430
Glu Glu Leu Val Gln Ile Cys Lys Lys Lys Lys Asp Glu Gln
435 440 445