卵巢上皮性癌SEREX抗原基因TM4SF1、C1D、TIZ原核表达载体的构建及表达  

CONSTRUCTION AND EXPRESSION OF PROKARYOTIC EXPRESSION VECTORS OF EPITHELIAL OVARIAN CANCER SEREX ANTIGEN GENES: TM4SF1、C1D、TIZ

作　　者：阳志军[1] 李力[1] 蒋燕明[1] 张玮[1] 张洁清[1] 潘忠勉[1] 

机构地区：[1]广西医科大学附属肿瘤医院妇瘤科,南宁530021

出　　处：《广西医科大学学报》2009年第3期362-365,共4页Journal of Guangxi Medical University

基　　金：广西科学研究与技术开发计划(No.桂科基0639043)

摘　　要：目的:克隆并构建卵巢上皮性癌SEREX抗原基因:TM4SF1、C1D、TIZ的原核表达载体。方法:用PCR从含SEREX抗原基因序列的重组pBK-CMV质粒中扩增TM4SF1、C1D、TIZ的编码序列(coding sequence,CDS),双酶切后插入原核表达质粒pET-30b(+),经酶切及测序鉴定,阳性质粒转化表达菌BL21感受态细胞,用IPTG诱导表达目的蛋白,用SDS-PAGE分析表达产物。结果:成功扩增了TM4SF1、C1D、TIZ的CDS,并构建了原核表达质粒pET-30b(+)/TM4SF1、pET-30b(+)/C1D、pET-30b(+)/TIZ,经测序与Genbank中的相应序列一致。并在大肠杆菌BL21中成功诱导表达目的蛋白。结论:成功构建了原核表达质粒pET-30b(+)/TM4SF1、pET-30b(+)/C1D、pET-30b(+)/TIZ,并在大肠杆菌中表达了重组融合目的蛋白,为进一步研究奠定了基础。Objective：The goal of the study was to amplify CDSes of epithelial ovarian cancer SEREX antigen genes：TM4SF1,C1D,TIZ and construct their prokaryotic expressing vectors. Methods：CDSes of TM4SF1, C1D,TIZ were amplified by PCR with plasmids DNA ,recombination pBK-CMV plasmids, that containing sequences of TM4SF1 ,C1D,TIZ and were inserted into pET-30b（＋） vector. Recombination plasmids were identified by restriction endonuclease analysis and DNA sequencing. Positive plasmids were transformed into E. coli BL21（DE3） for expression recombination protein under induction of IPTG. The expressed products were identified by SDS-PAGE. Results： CDSes of TM4SF1, C1D and TIZ were amplified successfully and cloned into pET-30b （＋） vectors. Sequences of these CDSes were confirmed by blasting to Genbank.Recombination proteins were successfully expressed in E. coli. Conclusion： Recombination pET- 30b（＋）/ TM4SFI,pET-30b（＋）/CID,pET-30b（＋）/ TIZ expression vectors were constructed and expressed successfully, which may provide basis for the further study of these antigen genes.

关 键 词：卵巢上皮性癌 SEREX 抗原基因 基因重组 

分 类 号：R737.31[医药卫生—肿瘤]