人BimS蛋白的原核表达及纯化  

Prokaryotic Expression and Purification of Human BimS Protein

作　　者：何侃[1] 佟立全[1] 战卓[1] 金英花[1] 

机构地区：[1]吉林大学分子酶学工程教育部重点实验室,长春130021

出　　处：《中国生物制品学杂志》2009年第9期880-881,886,共3页Chinese Journal of Biologicals

基　　金：国家自然科学基金(30770477)

摘　　要：目的克隆人BimS基因,原核表达并纯化目的蛋白。方法用PCR方法从HeLa细胞cDNA文库中扩增人BimS基因,克隆至pGEX-6P-1表达载体,转化大肠杆菌BL21(DE3),IPTG诱导表达。表达产物经GST亲和层析纯化。结果PCR扩增得到327bp的DNA片段,重组表达质粒pGEX-6P-1-BimS经双酶切鉴定和测序分析,表明构建正确。表达的目的蛋白相对分子质量约37000,表达量约占菌体总蛋白的30%,纯化后纯度可达93%。结论已成功克隆并原核表达了人BimS基因,得到纯度较高的BimS蛋白,为进一步研究其结构和功能奠定了基础。Objective To clone human BimS gene, expressed in prokaryotic cells and purify the expressed protein. Methods Human BimS gene was amplified from eDNA library of HeLa cells by PCR and cloned into expression vector pGEX-6P-1. The construeted recombinant plasmid pGEX-6P-1-BimS was transformed to E. coli BL21 （DE3） for expression under induction of IPTG. The expressed product was purified by GST affinity chromatography. Results A DNA fragment at length of 327 bp was amplified by PCR. Both restriction analysis and sequencing proved that recombinant plasmid pGEX-6P-1-BimS was constructed correctly. The expressed product, with a relative molecular mass of about 37 000, contained 30% of total somatic protein and reached a purity of 93% after purification. Conclusion Human BimS gene was successfully cloned and expressed in prokaryotic cells, and BimS protein with high purity was obtained, which laid a foundation of further study on structure and function of BimS protein.

关 键 词：人BimS基因 克隆 原核表达 纯化 

分 类 号：R373.2[医药卫生—病原生物学] R392.11[医药卫生—基础医学]