白皮黄瓜鲨烯合酶基因cDNA克隆及生物信息学分析  

Cloning and Bioinformatic Analysis of Squalene Synthase Gene from White Cucumber

作　　者：马成通 钱洁颖 刘镛[3] 陶晨陈 苏荷玲 晁耐霞[1,2] 吴耀生[1,2] 

机构地区：[1]广西医科大学生物化学与分子生物学教研室,南宁530021 [2]广西高校生物分子医学研究重点实验室,南宁530021 [3]广东医学院药学院,东莞523808 [4]桂林医学院,桂林541004

出　　处：《基因组学与应用生物学》2016年第1期183-189,共7页Genomics and Applied Biology

基　　金：国家自然科学基金(No.31260069);广西高等学校科研项目(No.201203YB038)共同资助

摘　　要：为认识葫芦科植物中葫芦素生物合成途径所需鲨烯合酶基因结构特征,克隆白皮黄瓜鲨烯合酶(squalene synthase,SS)基因c DNA并进行生物信息学分析。根据葫芦科植物绞股蓝、罗汉果、红花栝楼等的鲨烯合酶基因cDNA序列,设计白皮黄瓜鲨烯合酶引物,分别采用3'RACE和5'RACE技术扩增鲨烯合酶基因3'端和5'端。获得白皮黄瓜鲨烯合酶基因的两个cDNA克隆,命名为CsSS1和CsSS2,其cDNA序列全长分别为1 627 bp和1 534 bp,都编码417个氨基酸残基,分子质量为47.6 k D。成功克隆得到白皮黄瓜鲨烯合酶基因全长cDNA序列并对其进行序列分析,后续可用于葫芦素生物合成途径所需鲨烯合酶基因正选择位点功能分析。We cloned the full length cDNA encoding squalene synthase （SS） from white cucumber and did bioi- nformatic analysis, in order to get more insight into the structural characteristics of SS involved in cucurbitacine biosynthesis pathway of Cucurbitaceae. According to the cDNA sequence of SS gene from Cucurbitaceae including Gynostemma pentaphyllum, Siraitia grosvenorii, Trichosanthes rubriflos and so on, primers of squalene synthase of white Cucumber were designed to amplify 3＇-terminal and 5＇-terminal sequence of SS by 3＇RACE and 5＇RACE respectively. Two cDNAs of SS of white cucumber were obtained, named CsSS1 （1 627 bp） and CsSS2 （1 534 bp） respectively, all coding a polypeptide containing 417 amino acid residues with molecular weight of 47.6 kD. We successfully cloned the full length eDNA sequence of white cucumber squalene synthase gene and did bioinformatic analysis. Furthermore, these two cDNA clones will be utilized for the analysis of the positive selection site of squalene synthase involved in cucurbitacine biosynthesis nathway.

关 键 词：白皮黄瓜 鲨烯合酶 RACE 基因克隆 生物信息学分析 

分 类 号：Q943.2[生物学—植物学]