猪表皮生长因子在大肠杆菌中的表达  

Expression of Porcine Epidermal Growth Factor in Escherichia coli

作　　者：杨桂香[1] 黄伟华[1] 彭险峰[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《华南农业大学学报》2006年第4期90-93,共4页Journal of South China Agricultural University

摘　　要：采用RT-PCR方法从猪肾脏皮质组织中扩增出pEGF基因,将其连接到克隆载体pMD18-T上,经测序鉴定正确后,再克隆到原核表达载体pET-41 a(+)上,构建重组表达质粒pET-EGF.用重组质粒转化E.coliBL21(DE3)宿主菌,经IPTG诱导表达后利用SDS-PAGE电泳和W estern-b lotting对GST-EGF融合蛋白进行分析,结果表明37℃诱导表达的融合蛋白含量较高,占菌体总蛋白的20.2%;30℃诱导表达的可溶性融合蛋白含量较高,占菌体总蛋白的7.7%.Porcine epidermal growth factor （pEGF） cDNA was amplified from the RNA of swine kidney cortex by RT-PCR, and then the amplicon was cloned into pMD18-T for sequencing. After confirmation by sequencing, the pEGF cDNA fragment was inserted into prokaryotic expression vector pET-41a（ ＋ ） to construct recombinant plasmid pET-EGF. Then the constructed plasmid was transformed into host cell E. coli BI21 （ DE3 ）, and induced with IPTG. The expression of GST-EGF fusion protein was analyzed by SDS-PAGE and Western-blotting. The SDS-PAGE result indicated that fusion protein GST-EGF was 20. 2% of the total bacterial proteins, and the yield of the soluble target protein accounted for 7.7% of the total bacterial proteins.

关 键 词：猪表皮生长因子 原核表达 Westem-blotting 

分 类 号：S858.28[农业科学—临床兽医学]