重组人Stathmin基因的克隆及表达  

Cloning and Expression of Recombinant Human Stathmin Gene

作　　者：陈越[1] 李晶华[2] 何侃[1] 金英花[1] 

机构地区：[1]吉林大学生命科学学院疫苗研究中心,长春130012 [2]吉林大学公共卫生学院,长春130021

出　　处：《中国生物制品学杂志》2006年第2期113-115,共3页Chinese Journal of Biologicals

基　　金：国家自然科学基金面向项目(30370294).

摘　　要：目的克隆人Stathmin基因,并利用原核细胞进行表达。方法用PCR方法从Hela细胞cDNA文库中扩增Stathmin基因,PCR产物经TA克隆并测序后,克隆至pGEX-4T-1载体,并转化至大肠杆菌BL21感受态细胞中,IPTG诱导表达,经SDS-PAGE和Westernblot鉴定。结果PCR扩增得到460bp的cDNA片段,经测序分析,与GenBank数据库报道的人Stathmin(NM_203401)序列一致,经SDS-PAGE和Westernblot证实诱导表达的蛋白为GST融合蛋白。结论利用原核表达系统表达了人Stathmin,为进一步研究Stathmin结构和功能奠定了基础。Objective To clone human stathmin gene and express in prokaryotic cells. Methods Stathmin gene was amplified from the cDNA library of Hela cells by PCR. After TA cloning and sequencing,the amplified gene was cloned into vector pGEX-4T-1 and transformed to competent E. coli BL21 for expression under induction of IPTG. The expressed product was identified by SDS-PAGE and Western blot. Results A 460 bp cDNA fragment was amplified by PCR, and its sequence was consistent with that of human stathmin （NM_203401） reported in C, enBank. The expressed product was proved as GST fusion protein by SDS-PAGE and Western blot. Conclusion Human stathmin was expressed in prokaryotic cells. It laid a foundation of further study on the structure and function of stathmin.

关 键 词：STATHMIN 克隆 原核表达 重组人Stathmin基因 

分 类 号：R394[医药卫生—医学遗传学]