大豆过氧化物酶基因的克隆及其表达载体的构建  

Cloning and Plasmid Construction of Soybean Peroxidase Gene for Prokaryotic Expression System

作　　者：曾家豫[1] 袁红霞[1] 廖世奇[2] 周思彤[1] 梁琼[1] 

机构地区：[1]西北师范大学生命科学学院,甘肃兰州730070 [2]甘肃省医学科学研究院,甘肃兰州730050

出　　处：《食品科学》2010年第5期197-200,共4页Food Science

基　　金：甘肃省教育厅科研项目(0601-28;0501B-16);甘肃省自然科学研究基金计划项目(096RJZA035)

摘　　要：大豆过氧化物酶(soy bean peroxidase,sbp)基因的克隆及其表达载体的构建将有利于人们更深入的从分子生物学角度研究sbp基因的结构与功能。首先从大豆根中获取总RNA,通过RT-PCR获得sbp基因。再将sbp基因与表达载体pPICZα-A双酶切后连接,构建重组表达载体pPICZα-A-sbp。将pPICZα-A-sbp转化大肠杆菌筛选阳性克隆体并测序。结果表明:获得的sbp基因序列与已报道的sbp[U51191(GmEpa1)]基因序列有92%的同源性。重组表达载体pPICZα-A-sbp的成功构建为其在毕赤酵母中表达奠定了基础。The cloning and plasmid construction of soybean peroxidase （sbp） for prokaryotic expression system will provide more information to study the structure and function of soybean peroxidase by means of molecular biology. Total RNA was extracted from soybean root. Soybean peroxidase gene was obtained by RT-PCR technique and transferred into pPICZα-A vector through restriction endonuclease digestion. The resulting recombinant pPICZα-A-sbp expression vector was transformed into E. coli and the amplified recombinant pPICZα-A-sbp expression vector was selected and sequenced. Results indicated that cloned DNA sequence from soybean root exhibited 92% homology to the reported sbp （U51191 （GmEpa1） DNA sequence. Therefore, pPICZα-A-sbp was successfully constructed, which will provide base for its expression in Pichia pastoris （Gs115）.

关 键 词：大豆过氧化物酶 基因克隆 表达载体 

分 类 号：Q785[生物学—分子生物学]