猪防御素基因PBD-I在大肠杆菌中的重组和融合表达

Recombination and Fusion Expression of Porcine Defensin Gene PBD-I in E.coli

用RT-PCR方法获得PBD I基因,将该基因插入原核表达载体PinPointTMXa-3中,重组质粒转化大肠杆菌JM109,经IPTG诱导后以融合蛋白形式表达。SDS-PAGE电泳显示PBD I基因在目标位置17kDa处获得了表达。PBD I基因的成功表达为进一步研究其抗菌活性、抗菌机理及应用研究形成基础。

The porcine defensin gene PBDI was amplified by RT-PCR,then the gene was inserted into expression vector PinPointTMXa-3.Recombinant plasmid named as ppd1 was transformed into E.Coli JM109,which could effectively produce fusion protein induced with IPTG.The positive clone of PBDI gene expressed 17kDa fusion protein by SDS-PAGE electrophoresis.Expression of PBDI gene didn't increase distinctly along with time.The expression of PBDI gene lays a foundation in research on antimicrobial activities and its mechanism of the defensin.