猪链球菌2型毒力因子gapdh基因的克隆·表达及与Hela细胞互作蛋白的筛选  

Cloning,Expression of Streptococcus suis Type 2 Virulence Factors gapdh gene and Screening for Protein with Hela Cell

作　　者：袁芳艳[1,2] 谭臣[2,3] 雷丽[3] 贝为成[2,3] 刘泽文[1] 郭锐[1] 段正赢[1] 杨克礼[1] 周丹娜[1] 田永祥[1] 

机构地区：[1]湖北省农业科学院畜牧兽医研究所/动物胚胎工程及分子育种湖北省重点实验室,湖北武汉430064 [2]农业微生物学国家重点实验室,湖北武汉430070 [3]华中农业大学动物医学院,湖北武汉430070

出　　处：《安徽农业科学》2014年第36期12821-12824,共4页Journal of Anhui Agricultural Sciences

摘　　要：[目的]克隆并在大肠杆菌中表达猪链球菌2型毒力因子gapdh基因,寻找与GAPDH互作的受体蛋白。[方法]扩增猪链球菌2型甘油醛-3-磷酸脱氢酶gapdh基因并构建表达质粒p ET28a-gapdh。[结果]在大肠杆菌BL21(DE3)中成功表达了约41 k Da的融合蛋白GAPDH。Western blot试验说明GAPDH具有良好的免疫学活性。利用新西兰大白兔制备抗GAPDH蛋白的多克隆抗体。运用配体重叠技术,在He La细胞中初步找到了一个与GAPDH蛋白作用的潜在受体蛋白。[结论]为进一步研究猪链球菌2型的分子致病机理奠定基础。[ Objective ] The aim was to study cloning, expression of streptococcus suis type 2 virulence factors gapdh gene and screening forprotein with Hela ceil.[Method ] The glyceraldehyde 3 - phosphate dehydrogenase gapdh gene of Streptococcus suis serotype 2 （S. suis 2, SS2 ）was amplified by PCR, and inserted into the expression plasmid pET28a. [ Result] About 41 kDa fusion protein was successfully expressed inescherichia coli BL21 （DE3）. Good immunological activity of GAPDH protein was confirmed by western blot test. The polyclong antibodies ofrabbit anti GAPDH protein was preparated. Using ligand overlapping technology, a possible GAPDH protein receptor proteins wag preliminarylyfound in HeLa . [ Conclusion] The study laid a foundation for further insights into the molecular pathogenesis of SS2.

关 键 词：2型猪链球菌 原核表达 多抗制备 受体 

分 类 号：S852.6[农业科学—基础兽医学]