山羊GSDM家族相关基因的克隆及细胞焦亡效应蛋白GSDMD-NT的原核表达  

Cloning of GSDM family related genes and prokaryotic expression of pyroptosis effector protein GSDMD-NT of Hainan black goat

作　　者：任旭皎 黎振标 刘健新[1] 张凯照 鲁荣 李慧子 张彭涛 宁章勇[1] REN Xu-jiao;LI Zhen-biao;LIU Jian-xin;ZHANG Kai-zhao;LU Rong;LI Hui-zi;ZHANG Peng-tao;NING Zhang-yong(College of Veterinary Medicine,South China A gric uhural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院

出　　处：《中国兽医科学》2018年第8期1043-1049,共7页Chinese Veterinary Science

基　　金：广东省科技计划项目(2016A020210079)

摘　　要：为了解GSDM家族蛋白在山羊疾病发生过程中的作用,采用RT-PCR方法克隆了海南黑山羊GSDM家族相关基因,并对其进行序列及编码蛋白结构分析;基于对GSDMD的氨基酸序列分析,寻找到了其Caspases切割位点,扩增了GSDMD-NT序列,并将其克隆至原核表达载体进行诱导表达。结果显示,海南黑山羊GSDM家族基因种间同源性低,都存在Gasdermin超家族结构域,是结构保守蛋白;His标签GSDMD-NT的原核表达质粒经诱导后表达出51.2 ku的融合蛋白,该蛋白可以与抗His标签的单克隆抗体发生特异性反应。本研究结果为GSDM家族蛋白功能的物种间比较研究提供了基础数据,也为进一步探究GSDMD-NT诱导细胞焦亡的机制奠定了基础。To understand the role of GSDM family proteins in the pathogenesis of goat diseases,the GSDM family related genes of Hainan black goat were cloned by RT-PCR,and their sequences and coding protein structures were analyzed. Based on the amino acid sequences of GSDMD,its caspases cleavage site was found,and the GSDMD-NT gene sequence was amplified and cloned into the prokaryotic expression vector for expression.In result,the GSDM family genes of Hainan black goat had low homology between species,and the encoded proteins were conservative containing Gasdermin superfamily structure domain. The prokaryotic expression plasmid of His-tag GSDMD-NT was induced to produce a fusion protein of51.2 ku in mass,and the fusion protein had specific reaction with anti-His monoclonal antibodies.The results provided the basic data for the comparative studies on the function of GSDM family proteins,and also laid a foundation for further exploration of GSDMD-NT induced pyroptosis.

关 键 词：海南黑山羊 GSDM家族基因 GSDMD-NT 原核表达 

分 类 号：S852.42[农业科学—基础兽医学]