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

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pET28a-gapdh.结果]在大肠杆菌BL21 （DE3）中成功表达了约41 kDa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HeLa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.

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

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.