人受精相关抗原-1的基因克隆及原核表达  

Cloning and expression of recombinant human fertilization antigen-1 in E. coli.

作　　者：张宏斌[1] 武婕[1] 王捷[1] 周晓宁[2] 

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

出　　处：《中国优生与遗传杂志》2009年第5期32-33,F0003,共3页Chinese Journal of Birth Health & Heredity

基　　金：广东省科技计划项目(2006C104004)

摘　　要：目的为了阐明免疫性不孕不育的原因,检测血清中存在的抗精子抗体的种类,构建重组人受精相关抗原-1(recombinant human Fertilization antigen-1,rhFA-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FA-1的cDNA,将其克隆入表达载体pBV220,构建人源性FA-1的重组原核表达质粒pBV220/FA-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FA-1基因完全一致。SDS-PAGE电泳显示,表达产物的相对分子量为14.6KDa与预期结果相符。结论获得了人FA-1的编码基因,并在大肠杆菌中表达了人的FA-1蛋白。Objective： To determine the anti - sperm antibody in serum of infertility patients, a recombinant prokaryotic expression plasmid of human fertilization antigen - 1 was constructed and the FA - 1 protein was expressed in E. coli. Methods： The code gene of human FA - 1 was amplified by reverse transcript PCR from human testes organism and cloned into a prokaryotic expression vector pBV220. The recombinant plasmid was identified by restriction enzyme digestion and DNA sequencing. The recombinant plasmid was transfected into E. coli JM109, then the recombinant protein was expressed in E. coli and detected by using SDS -PAGE. Results ： The sequence of recombinant human FA - 1 was correct. The relative molecular mass of the expressed product was identical to that of expectation of 15KDa. Conclusion： The code gene of human FA- 1 has been successfully cloned and expressed in E. coli.

关 键 词：rhFA-1 基因克隆 原核表达 

分 类 号：R392.11[医药卫生—免疫学]