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邵珠卿, 韩明月, 葛俊超, 王长军, 潘秀珍, 唐家琪. 猪链球菌2型组氨酸三聚体蛋白原核表达[J]. 中国公共卫生, 2009, 25(6): 669-671. DOI: 10.11847/zgggws2009-25-06-14
引用本文: 邵珠卿, 韩明月, 葛俊超, 王长军, 潘秀珍, 唐家琪. 猪链球菌2型组氨酸三聚体蛋白原核表达[J]. 中国公共卫生, 2009, 25(6): 669-671. DOI: 10.11847/zgggws2009-25-06-14
SHAO Zhu-qing, HAN Ming-yue, GE Jun-chao, . Prokaryotic expression of histidine triad protein from Streptococcus suis 2[J]. Chinese Journal of Public Health, 2009, 25(6): 669-671. DOI: 10.11847/zgggws2009-25-06-14
Citation: SHAO Zhu-qing, HAN Ming-yue, GE Jun-chao, . Prokaryotic expression of histidine triad protein from Streptococcus suis 2[J]. Chinese Journal of Public Health, 2009, 25(6): 669-671. DOI: 10.11847/zgggws2009-25-06-14

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

  • 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。

     

    Abstract: Objective To identify histidine triad prote in(HTP)gene in Streptococcus suis sero type 2(S.suis 2),and to study its an tigenicity.Methods Bioinformatics was adopted to analyze the genome database of the Chinese stra in 05ZYH 33 of S.suis 2.Prmiers were designed for HTP,and the target DNA fragment was amplified using the genomic template o f 05ZYH33.HTP gene was subsequently inserted into pEASY-T1 vector and subcloned into expression vector pET28a.The plasmid was confirmed by direc tDNA sequencing and tran sformed in to E.coli BL21(DE3)competent cells to express recom binant HTP.The recom binant prote in was purified with His-Resin.We stern blot was used to identify the antigenicity of recom binant HTP.Results The 2.7kbhtp sequence was amplified from the genome of 05ZYH 33,and the recombinant HTP can be produced in E.coli.A 110 kD prote in can be observed in SDS-PAGE,and Western blot experiment demon strated that the prote in 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.

     

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