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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 Pro-tein (HTP) of Streptococcus suis serotype 2 (Streptococcus suis serotype 2) and study the immunogenicity of this protein. Lay the foundation for the experiment. Methods The HTP gene was found in the whole genome of Streptococcus suis serotype ZZZZH33 by comparing with related family proteins. The synthetic primers were designed and amplified by PCR. The target fragment was cloned into the expression vector pET28a, expressed and purified, and its immunogenicity was detected by Western blot. Results The fragment of about 2.7kb was amplified by PCR and the target protein of about 110KD was successfully expressed. By His-Tag affinity chromatography, to obtain a higher purity of the fusion protein. Western blot results showed that the expressed product was immunogenic. Conclusions The Hsu-2 strain is highly virulent to S.suis2 and it has good immunogenicity and can be used as a candidate vaccine for S.suis2 immunoprotective vaccine.