兔血清对氧磷酶基因的克隆及其在大肠杆菌中的表达纯化  

Cloning,expression and purification of rabbit serum paraoxonase in Escherichia coli

作　　者：谷立坤[1] 张建云[1] 

机构地区：[1]河南工程学院资源与环境工程系,河南郑州451191

出　　处：《中国酿造》2011年第8期46-49,共4页China Brewing

基　　金：河南省科技攻关项目(092102210212);河南工程学院工程技术研究中心建设项目资助(2009ETRCHNIE02)

摘　　要：利用RT-PCR扩增新西兰大白兔肝脏PON1编码区全长序列,克隆入表达载体pET32a中,构建了重组表达载体pET32a-PON1,转化大肠杆菌获得表达菌株。SDS-PAGE表明该重组表达蛋白具有部分可溶性,酶活测定结果显示该可溶性蛋白具有降解芳香酯酶和内酯酶的活性。该研究实现了兔血清对氧磷酶在大肠杆菌中的功能性表达,并获得了纯化的蛋白,为下一步对氧磷酶PON1的功能研究奠定基础。The paraoxonase gene PON1 was cloned from rabbit liver by RT-PCR, and cloned into PET32a vector and constructed the PET32a-PON1 recombinant plasmid, wased into component cells of and the Expression strain was obtained by transformation of PET32a-PON1 plasmid into E. coli. SDS-PAGE analysis revealed that the recombinant proteins could be partially soluble. Enzyme assay indicated that the recombinant proteins had arylestemse and lactonase activities. The results made a good base for studying the functions of purified PON1.

关 键 词：逆转录PCR 原核表达 对氧磷酶 可溶性蛋白 

分 类 号：Q93-33[生物学—微生物学]