中文摘要：

为了验证双向电泳筛选出的7型猪链球菌WH分离株免疫相关蛋白乳酸脱氢酶的反应原性,本实验采用克隆表达的方法对其进行研究。根据质谱鉴定的NCBI登录号查找乳酸脱氢酶基因序列,并设计特异性引物,对乳酸脱氢酶基因进行扩增,克隆入p ET30a载体,构建重组质粒p ET30a-LDH,测序后转化入大肠埃希氏菌中。用IPTG进行诱导重组菌,SDS-PAGE和Western blot分析表达产物。SDS-PAGE分析结果显示表达的重组融合蛋白相对分子量为41ku;Western blot分析结果显示该重组蛋白可与7型猪链球菌WH分离株多克隆抗体发生特异性血清学反应。这表明表达的重组LDH蛋白具有良好的反应原性,为预防和控制猪链球菌病提供有效的疫苗候选分子奠定基础。

英文摘要：

To verify the reactogenici ty of immune-related protein lactate dehydrogenase （LDH） which was screened from Streptococcus sois type 7 WH isolate by two-dimensional electrophoresis.According to LDH gene sequences accessed in NCBI, a pair of specific primers was desigend to amplify LDH gene.The amplified product was cloned into expression vector pETB0a for expressing.Recombinant plasmid pET30a-LDH was transformed into E.coli.The expression products were analysed by SDS-PAGE and Western blot after IPTG induction.SDS-PAGE results confirmed that the recombinant fusion protein was expressed with a relative molecular weight of 41 ku;western blot analysis showed that the recombinant protein could specifically be reactive to S.suis type7 WH polyclonal antibody,which confirmed that the recombinant LDH protein was endowed with good reactionogenicity.The study provides an effective vaccine candidate molecule to prevent and control S.sois disease.