绵羊Granulysin在大肠杆菌中的表达与鉴定  

Expression and Identification of Ovine Granulysin in E.coli

作　　者：杨国宇[1] 乔新安[1] 朱彦彩[2] 贾海丽[1] 王月影[1] 韩立强[1] 王艳玲[1] 

机构地区：[1]河南农业大学动物生理生化实验室,河南郑州450002 [2]河南科技学院细胞工程重点实验室,河南新乡453003

出　　处：《江西农业学报》2007年第11期61-63,共3页Acta Agriculturae Jiangxi

基　　金：河南省重点科技攻关项目(编号:0523010500)

摘　　要：根据GenBank中公布的绵羊Granulysin基因序列设计引物,用PCR法从重组质粒中扩增出含BamHI/XhoI酶切位点的Granulysin片段,双酶切后克隆至pGEX-4T-1载体,构建重组原核表达质粒pGEX-Granulysin,转化宿主菌大肠杆菌BL21(DE3),经IPTG诱导进行表达,SDS-PAGE鉴定,表达的融合蛋白分子量约为41 kDa,并以包含体形式存在。The primers were designed according to the sequences of ovine Granulysin from GenBank. A fragment of ovine Granulysin containing BamHI/XhoI was amplified from recombinant plasmid by PCR. The fragment cleaved by BamHI/XhoI was subcloned into pGEX -4T - 1 vector and to construct a recombinant expression plasmid of pGEX - Granulysin, Subsequently, E. coli BL21 （DE3） competent cells were transformed by the recombinant plasmid. The fusion protein, which was induced by IPTG was identified by SDS - PAGE. The fusion protein expressed in E. coli was 41 kDa and mainly existed as inclusion bodies.

关 键 词：绵羊 颗粒溶解素 原核表达 鉴定 大肠杆菌 

分 类 号：S826[农业科学—畜牧学]