家蝇幼虫抗菌蛋白基因AS566在毕赤酵母中的表达及产物抑菌活性检测  

Expression of Musca domestica larva antibacterial substance gene AS566 in Pichia pastoris and its antibacterial activity

作　　者：闫恕[1] 李文婷[1] 丁士雯 袁野[1] 唐艳[1] 孔令聪[1] 裴志花[1] 刘树明[1] 马红霞[1,2] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]吉林农业大学动物生产及产品质量安全教育部重点实验室,吉林长春130118

出　　处：《中国兽医科学》2016年第3期344-347,共4页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31140026;31572574;31502121);吉林省世行贷款农产品质量安全项目(2011-Y05)

摘　　要：为采用巴斯德毕赤酵母系统表达家蝇幼虫抗菌蛋白AS566基因,并对其抑菌活性进行初步确证。以分子克隆法构建重组表达质粒pGAPZαA-AS566,将其线性化后电击转化至毕赤酵母GS115感受态细胞,在GAP启动子调控下,于30℃诱导表达,SDS-PAGE检测结果显示表达产物的分子质量约为43.5 ku,与预期结果一致。采用管碟法对表达产物的抑菌活性进行检测,结果显示该表达产物对鸡源沙门菌和猪源金黄色葡萄球菌的生长具有抑制作用。本研究为进一步研究家蝇幼虫抗菌蛋白的抑菌机制奠定了基础。To express Musca domestica larva antibacterial substance gene（AS566） in Pichia pastoris and also analyze the antibacterial activity initially, the recombinant expression plasmid p GAPZα-A-AS566 was constructed under Molecular Cloning Laboratory Manual, and electroporated in the P. pastoris GS115. AS566 gene was expressed at 30 ℃ under the control of GAP promoter,SDS-PAGE result showed that the molecular weight was about 43.5 ku as expected. The antibacterial activity was analyzed using a agar-diffusion method, the result showed that the recombinant expression plasmid p GAPZαA-AS566 was successfully expressed in the P. pastoris and could inhibit the growth of Salmonella from chicken and Staphylococcus aureus from swine. This study would lay the foundation for further study on antibacterial mechanism of M.domestica larva antibacterial substance.

关 键 词：家蝇抗菌蛋白(AS566) 毕赤酵母 真核表达 抑菌活性 

分 类 号：S852.743[农业科学—基础兽医学]