人卵透明带重组融合基因及其表达载体的构建  

Construction of recombination human zona peiiucida3/GST fusion protion gene expression vector

作　　者：郭焱[1] 路英丽[1] 邹冬辉[1] 赵慧[1] 王忠山[1] 

机构地区：[1]吉林大学基础医学院细胞生物学教研室,吉林长春130021

出　　处：《中国现代医学杂志》2006年第15期2265-2268,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金资助项目(39870313)

摘　　要：目的构建人卵透明带蛋白3(zonapellucida3,ZP3)/GST蛋白融合基因原核表达载体。方法利用PCR法扩增出去N端的信号肽和C端的跨膜区huZP3成熟蛋白编码基因。先将其克隆到pGEM-T载体,进行序列测定和分析,然后将huZP3核心DNA片断克隆入GST蛋白融合原核表达载体pGEX4T-1内。结果获得一个核苷酸长度约为1000bp的基因,与GenBank(NCBI:M60504)公布的ZP3基因序列有99%的同源性,DNA序列分析发现,有一个氨基酸出现变异。酶切鉴定后证明ZP3基因完整插入GST融合蛋白原核表达载体pGEX4T-1中。结论成功构建出含ZP3基因的GST融合蛋白原核表达载体。[Objective] To construct GST fusion protion gene expression vector which can express human zona peiiucida in Prokaryotic cells. [Methods] The ZP3 gene was amplified by using PCR. The amplified DNA fragment was cloned into pGEM-T vector for sequencing analysis. The ZP3 gene was then subcloned into fusion protion expression vector pGEX4T-1. [Result] A gene of 1 000 bp was obtained and showed a 99% homology with ZP3 gene sequence published in GenBank （NCBI： M60504）. ZP3 gene was inserted into Prokaryotic expression vector pGEX4T-1 correctly. [Conclusion] Having constructed ZP3 GST protion expression vector successfully.

关 键 词：人卵透明带蛋白3 GST融合蛋白 表达载体构建 

分 类 号：R392.11[医药卫生—免疫学]