人Bik基因的克隆、表达与纯化  

Cloning and Expression of Human Bik Gene and Purification of Expressed Product

作　　者：李晶华[1] 何侃[1] 佟立全[1] 李杨[1] 高畅[1] 孔维[1] 金英花[1] 

机构地区：[1]吉林大学生命科学院

出　　处：《中国生物制品学杂志》2008年第7期581-583,共3页Chinese Journal of Biologicals

基　　金：国家自然科学基金项目(30640064;30770477)

摘　　要：目的克隆人Bik基因,并进行表达及纯化。方法用PCR方法从HeLa细胞cDNA文库中扩增Bik基因,克隆至pGEX-6P-1表达载体,转化至大肠杆菌BL21(DE3)中,IPTG诱导表达。表达产物进行GST亲和层析纯化。结果PCR扩增得到483bp的DNA片段,重组质粒pGEX-6p-1-Bik经酶切鉴定和测序分析,表明构建正确。表达的重组蛋白占菌体总蛋白的38%,纯化后蛋白纯度达96%。结论已成功克隆并利用原核表达系统表达了人Bik基因,为进一步研究Bik蛋白结构和功能奠定了基础。Objective To clone and express human Bik gene and purify the expressed product. Methods Amplify Bik gene from the cDNA library of HeLa cells by PCR and clone into expression vector pGEX-6P-1. Transform the constructed recombinant plasmid pGEX-6P-1-Bik to E.coli BI21 （DE3） for expression under induction of IPTG. Purify the expressed product by GST affinity column chromatography. Results A DNA fragment at length of 483 bp was amplified. Both restriction analysis and sequencing proved that recombinant plasmid pGEX-6P-1-Bik was correctly constructed. The expressed product contained about 38% of total somatic protein and reached a purity of 96% after purification. Conclusion Human Bik gene was successfully cloned and expressed in prokaryotic cells, which laid a foundation of further study on structure and function of Bik protein.

关 键 词：Bik基因 克隆 原核表达 纯化 

分 类 号：Q785[生物学—分子生物学] Q786