猪链球菌2型组氨酸三聚体蛋白原核表达  

Prokaryotic expression of histidine triad protein from Streptococcus suis 2

作　　者：邵珠卿[1,2] 韩明月[1,2] 葛俊超[1,2] 王长军[2] 潘秀珍[1,2] 唐家琪[2] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210046 [2]南京军区军事医学研究所

出　　处：《中国公共卫生》2009年第6期669-671,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金(30730081,30600533,30670105);南京军区医学科技创新课题(072045);南京军区卫生专业人才培养“122”工程资助项目;江苏省自然科学基金资助项目(BK2008066,BK2007013)

摘　　要：目的鉴定猪链球菌2型(Streptococcus suis serotype2,S.suis2)组氨酸三聚体蛋白(Histidine Triad Pro-tein,HTP),并研究该蛋白的免疫原性,为研究S.suis2保护性疫苗奠定实验基础。方法通过与相关家族蛋白进行同源性比较,在猪链球菌Z型05ZYH33全基因组序列中发现了编码HTP的基因。设计合成引物,进行PCR扩增,并将目的片段克隆到表达载体pET28a,表达并纯化出目的蛋白,通过蛋白印迹(Western blot)检测其免疫原性。结果PCR扩增出约2.7kb的片段,并成功表达分子量在110KD左右的目的蛋白。通过His-Tag亲和层析,获得纯度较高的融合蛋白。Western blot检测结果表明,该表达产物具有免疫原性。结论S.suis2中国强毒株中存在HTP,并具有良好免疫原性,可作为S.suis2免疫保护性疫苗候选分子。Objective To identify histidine triad protein（HI＇P） gene in Streptococcus suis serotype 2 （S. suis 2 ）, and to study its antigenicity. Methods Bioinformatics was adopted to analyze the genome database of the Chinese strain 05ZYH33 of S. suis 2. Primers were designed for HTP,and the target DNA fragment was amplified using the genomic tem- plate of 05ZYH33. HTP gene was subsequently inserted into pEASY-T1 vector and subeloned into expression vector pET28a. The plasmid was confirmed by direct DNA sequencing and transformed into E. coli BL21 （ DE3 ） competent cells to express recombinant HTP. The recombinant protein was purified with His-Resin. Western blot was used to identify the antigenicity of recombinant HTP. Results The 2.7kb htp sequence was amplified from the genome of 05ZYH33, and the recombinant HTP can be produced in E. coil A 110 kD protein can be observed in SDS-PAGE, and Western blot experi- ment demonstrated that the protein shared strong specific antigenicity. Conclusion HTP can function as a novel antigen of S. suis 2, and may play pivotal roles in the severe infection of S. suis 2.

关 键 词：猪链球菌2型 组氨酸三聚体蛋白 克隆 表达 

分 类 号：R387.12[医药卫生—医学寄生虫学]