山葡萄UDP_葡萄糖_类黄酮_3_省略_因_3GT_cDNA的克隆和分析_刘海峰.pdfVIP

DOI:10.13592/ki.ppj.2009.08.008 74 8 植物生理学通讯 第45 卷第8 期，2009 年8 月 研究报告 Original Papers 山葡萄UDP- 葡萄糖:类黄酮-3-O- 葡萄糖基转移酶基因(3GT) cDNA 的克 隆和分析 1,2 1 1 1,* 刘海峰 ,杨成君,于淼,王军 1 东北林业大学林木遗传育种与生物技术教育部重点实验室, 哈尔滨150040; 2 延边大学农学院, 吉林龙井133400 提要: 用RT-PCR和SMART RACE技术克隆了山葡萄3GT基因的全长cDNA序列。GenBank登录号为FJ169463,3GT cDNA 全长1 477 bp,包括开放阅读框1 371 bp,编码456个氨基酸,该基因表达产物相对分子质量为50.19 kDa,等电点为5.98,是 不稳定蛋白。该基因属于UGT超基因家族,不包含信号肽。氨基酸序列与欧亚种葡萄(AB047098)、美洲种葡萄(EF630356) 和拟南芥(AY072325)等植物的3GT氨基酸序列的同源性系数分别为98%、97%和59%。 关键词:山葡萄; cDNA克隆; UDP-葡萄糖:类黄酮-3-O-葡萄糖基转移酶 cDNA Cloning and Analysis of UDP-Glucose:Flavonoid 3-O-Glucosyltransferase (3GT) in Vitis amurensis 1,2 1 1 1, * LIU Hai-Feng , YANG Cheng-Jun , YU Miao , WANG Jun 1K ey L aboratory of F orest Tree Genetic Imp rovement and Biotechnology, M inistry of Education, N ortheast F orestry University, 2 Harbin 150040, China; Ag ricultural College of Yanbian University, Longj ing, J ilin 133400, China Abstract : The full-length cDNA sequences of 3GT genes in Vitis amurensis were cloned by the techniques of RT-PCR and SMART RACE . The landing GenBank N o. is FJ 169463 and the full-length of 3GT cDNA is 1 477 bp with open reading frame of 1 37 1 bp, which encodes 456 amino acids. The product of 3GT gene expression is unstable protein with the molecular mass of 50.19 kDa, isoelectric point of 5.98. The 3GT gene belongs to the UGT supergene family, not containing a signal peptide. The amino sequence of 3GT in Vitis amurensis has the homology with 3GT of Vitis vinif era, Vitis labrusca and Arobidop sis thaliana, and the like- ness coefficient is 98%, 97% and 59% respecti