重组人睾丸特异性抗原-1(TSA-1)的基因克隆及原核表达  

Cloning and expression of recombinant human testis specific antigen-1 related to infertility in Ecoli

作　　者：周晓宁[1] 武婕[2] 张宏斌[2] 王捷[2] 

机构地区：[1]广州军区广州总医院妇产科,广东广州510010 [2]广州军区广州总医院医学实验科,广东广州510010

出　　处：《中国优生与遗传杂志》2008年第10期29-30,共2页Chinese Journal of Birth Health & Heredity

基　　金：广东省科技计划项目(2006C104004)

摘　　要：目的为了阐明血清中存在的抗精子抗体引起不孕不育的原因,构建重组人睾丸特异性抗原-1(recombinanthuman testis specific antigen-1,rhTSA-1)的原核表达质粒并在大肠杆菌中诱导表达。方法采用RT-PCR法,从人睾丸组织总RNA中扩增获得人TSA-1的cDNA,将其克隆入表达载体pGEX-4T-2,构建TSA-1的重组原核表达质粒pGEX/TSA-1。重组质粒经酶切和测序鉴定后,转化大肠杆菌JM109并在大肠杆菌中诱导表达目的蛋白。结果测序表明,重组基因序列与人TSA-1基因完全一致。SDS-PAGE电泳显示,表达产物的相对分子量与预期结果相符。结论获得了人TSA-1的编码基因,并在大肠杆菌中可表达人TSA-1。Objective:To determine the anti-sperm antibody in serum of infertility patients,a recombinant prokaryotic expression plasmid of human testis specific antigen-1 was construct and the protein was expressed in E.coli.Methods: The code gene of human TSA-1 was amplified from human testes organism and cloned into a prokaryotic expression vector pGEX-4T-2.The recombinant plasmid was identified by DNA sequencing.The recombinant plasmid was transfected into E.coli JM109,then the recombinant protein was expressed in ...

关 键 词：TSA-1 基因克隆 原核表达 

分 类 号：R392[医药卫生—免疫学]