论文部分内容阅读
利用原核表达载体pCold I构建肠炎沙门菌转录调控蛋白HilD和HilA的重组表达菌,并采用Western-blot技术检测两者在体外培养条件下的表达规律。结果表明:成功构建了重组表达大肠埃希菌(E.coli)BL21(pCold I-hilD)和E.coli BL21(pCold I-hilA),经IPTG诱导得到了重组表达的rHis-HilD和rHis-HilA蛋白。将重组表达的蛋白免疫BALB/c小鼠,制备蛋白的多克隆抗体,Western-blot进一步证明了抗体与蛋白可发生特异性反应。肠炎沙门菌C50041在LB培养条件下,HilD与HilA蛋白表现出不同的表达规律,从而为体外研究肠炎沙门菌SPI1与SPI2效应蛋白的表达和功能研究奠定了基础。
The prokaryotic expression vector pCold I was used to construct the recombinant expression strains of HilD and HilA of Salmonella enteritidis. Western-blotting was used to detect the expression of the two genes in vitro. The results showed that recombinant plasmid pCold I-hilD and pCold I-hilA were successfully constructed, and recombinant recombinant rHis-HilD and rHis- HilA protein. The recombinant protein was immunized BALB / c mice to prepare protein polyclonal antibodies, Western-blot further confirmed that the antibody and protein can react specifically. Under the conditions of LB culture, HilD and HilA proteins show different expression patterns, which laid the foundation for the study of the expression and function of Salmonella enteritidis SPI1 and SPI2 effector proteins in vitro.