粤北三黄鸡β-防御素基因的表达及其生物活性研究

Study on expression and biological activity of β-defensin gene of yellow—feathered broiler in northern Guangdong province

采用RT-PCR方法从三黄鸡肝脏中扩增β-防御素Gal-6基因的cDNA片段,构建重组表达质粒pGEX-6p-1-GAL-6,将重组质粒转化至大肠杆菌BL21进行原核表达。结果表明,成功获得201 bp的鸡β-防御素基因,SDS-PAGE电泳分析表明,原核表达的重组鸡融合蛋白分子质量约为32 kD。此外,表达的该重组蛋白对金黄色葡萄球菌有较高抗菌活性。

β-defensin gallinacin-6 （Gal-6）cDNA fragment was amplified from liver of yellow-feathered broiler by RT-PCR.Sequence analysis of β-defensin showed that the gene was consiststed of 201 bp.β-defensin gene was cloned in-to pGEX-6p-1 vector and transfered into E.coli cells.SDS-PAGE analysis showed that a 32 kD protein equivalent to chicken protein in molecular weight was expressed in E.coli.The antimicrobial activity of the recombinant fusion protein was evaluated in vitro .which exhibited high antimierobial activity against Staphyloccocus aureus.