绵羊Granulysin在大肠杆菌中的表达与鉴定

Expression and Identification of Ovine Granulysin in E.coli

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.