膜荚黄芪UCGT基因的克隆及表达分析  

Cloning and expression analysis of UCGT genein Astragalus membranaceus

作　　者：赵洋 李俊杰 苏喆莹 李子羊 全雪丽[1] 吴松权[1] ZHAO Yang;LI Junjie;SU Zheying;LI Ziyang;QUAN Xueli;WU Songquan(Agricultural College of Yanbian University,Yanji Jilin 133002,China;Biotech Pharmaceutical CO.,LTD,Beijing 100176,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]百泰生物药业有限公司,北京100176

出　　处：《延边大学农学学报》2021年第3期1-7,共7页Agricultural Science Journal of Yanbian University

基　　金：国家自然科学基金资助项目(21462044);国家自然科学基金资助项目(30860036);吉林省自然科学基金资助项目(201115228,202101ZYTS197)。

摘　　要：UDP-毛蕊异黄酮葡萄糖基转移酶(UDP-glucose:calycosin-7-O-glucosyltransferase,UCGT)是糖基化毛蕊异黄酮生物合成毛蕊异黄酮葡萄糖苷的一种酶。试验采用同源克隆法和RACE技术从膜荚黄芪中克隆得到AmUCGT基因,对其进行生物信息学分析,同时调查了AmUCGT在不同组织中的表达量与毛蕊异黄酮葡萄糖苷含量之间的关系。结果表明:AmUCGT基因的全长cDNA序列长度为1862 bp,其中,开放阅读框(ORF)1455 bp,预测其编码484个氨基酸多肽,蛋白质分子量约为53.99 kDa,理论等电点pI为6.52,为可溶性蛋白。AmUCGT在根、茎和叶中的表达趋势与毛蕊异黄酮葡萄糖苷的含量变化一致,推测其在毛蕊异黄酮葡萄糖苷的生物合成中起着重要作用。UDP-glucose:calycosin-7-O-glucosyltransferase(UCGT)is an enzyme for biosynthesis of calycosin-7-O-β-D-glucoside by glycosylating calyconsin.In this study,AmUCGT gene was cloned from Astragalus membranaceus by using the homologous cloning and RACE technologies and its bioinformatic analysis was performed.Furthermore,the relationship between AmUCGT expression and calycosin-7-O-β-D-glucoside content in different organs of A.membranaceus was investigated.The results showed that the full cDNA length of AmUCGT was 1,862 bp and the open reading frame was 1,455 bp;AmUCGT was predicted to encode 484 amino acid polypeptides,its molecular weight was approximately 53.99 kDa,and the theoretical isoelectric point was 6.52.AmUCGT was a soluble protein.The change of AmUCGT expression was consistent with that of the calycosin-7-O-β-D-glucoside content in roots,stems and leaves of A.membranaceus,speculating that AmUCGT played an important role in biosynthesis of the calycosin-7-O-β-D-glucoside.

关 键 词：膜荚黄芪 UCGT 基因克隆 表达分析 

分 类 号：S567.239[农业科学—中草药栽培]