论文部分内容阅读
目的:从猪链球菌05ZY中扩增SSU1664基因,在大肠杆菌中表达并评价其重组蛋白的活性。方法:根据05ZY基因组序列设计引物,PCR扩增SSU1664基因,构建表达载体,利用亲和层析对目的蛋白进行纯化,Western blot和ELISA法鉴定其免疫原性。结果:SSU1664基因能够在大肠杆菌中可溶性表达,Western blot显示其能与猪链球菌菌体免疫后的大鼠血清反应,ELISA检测显示在人感染猪链球菌患者血清中针对SSU1664蛋白的IgM含量显著性升高。结论:SSU1664蛋白具有较高的免疫原性,可作为潜在的猪链球菌疫苗候选分子和猪链球菌早期感染诊断的标志物。
Objective: To amplify SSU1664 gene from Streptococcus suis 05ZY and express it in E. coli and evaluate the activity of its recombinant protein. Methods: Primers were designed based on the 05ZY genomic sequence. The SSU1664 gene was amplified by PCR. The expression vector was constructed. The target protein was purified by affinity chromatography. The immunogenicity was identified by Western blot and ELISA. Results: The gene of SSU1664 was soluble in E.coli and Western blot showed that it could react with the serum of S. suis cells after immunization. The results of ELISA showed that the IgM content of SSU1664 protein in serum of patients infected with Streptococcus suis Sexually elevated. CONCLUSIONS: SSU1664 protein has high immunogenicity and can be used as a potential diagnostic marker of Streptococcus suis for potential candidate Streptococcus suis vaccine.