人era基因的克隆测序及表达  

Cloning, sequencing and expression of cDNA gene of human era

作　　者：吴元明[1] 张俊杰[1] 纪宗玲[1] 刘惠萍[1] 陈南春[1] 陈苏民[1] 

机构地区：[1]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2000年第5期646-648,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金资助项目!(39870380;39670006);全军医药卫生科研基金资助项目!(98M108)

摘　　要：目的　克隆人的 era基因 (简称 Hera)并利用大肠杆菌进行表达 .方法　 PCR扩增 Hera基因 ,测序正确后克隆入大肠杆菌融合表达载体 p GEX- 4T3,受控于 Ptac启动子 ,重组质粒 p GEX- Hera以大肠杆菌 DH5α为宿主菌 ,用 IPTG进行诱导表达 .结果　克隆了 Hera基因 ,测序正确 ;含重组质粒p GEX- Hera的菌体诱导后在 SDS- PAGE上出现一条新生蛋白带 ,相对分子质量为 6 5 ku,占菌体总蛋白的 2 3% .结论　成功扩增、克隆 Hera基因 。AIM To amplify and to clone the human era gene and to express its recombinant protein in E.coli. METHODS After the human era gene was amplified by PCR and identified by sequencing, it was inserted into expression vector pGEX4T3 in which exogenous gene was controlled by ptac promoters. The recombinant plasmid pGEXHera was transformed into DH5α and induced with IPTG chemically. RESULTS The human and sequenced correctly. When the engineered bacteria were induced, an anticipated 65 ku protein band appeared on SDSnted to 23% of total bacterial protein. CONCLUSION The human era gene has been successfully subcloned and efficiently expressed in E.coli.

关 键 词：ERA基因 基因克隆 序列测定 表达 大肠杆菌 

分 类 号：Q78[生物学—分子生物学]