鸡肠炎沙门氏菌外膜蛋白F的原核表达及免疫原性分析

Prokaryotic Expression and Immunogenicity Analysis of Outer Membrane Protein F of ChickenSalmonella enteritidis

根据GenBank中收录的鸡肠炎沙门氏菌OmpF基因序列设计1对引物,以鸡肠炎沙门氏菌辽宁分离株基因组为模板,扩增鸡肠炎沙门氏菌OmpF基因,用IEDB Analysis Resource在线预测分析,该基因可能存在15个线性B细胞抗原表位;同时将该基因克隆至原核表达载体PGEX-4T-1中,构建重组表达质粒PGEX-4T-1-OmpF,转入大肠杆菌BL21中,获得OmpF重组蛋白。Western-blot检测结果表明,该重组蛋白的分子质量约为66 kD(含标签蛋白),能被鸡沙门氏菌阳性血清识别,初步证实该蛋白具有免疫原性,为鸡肠炎沙门氏菌亚单位疫苗的进一步研究和开发奠定了理论基础。

According to the sequence of OmpF gene of chicken Salmonella enteritidis collected in GenBank, a pair of primers were designed to amplify the OmpF gene of chicken Salmonella enteritidis Liaoning isolate with the genome as template. The gene was predicted and analyzed online by IEDB Analysis Resource, and 15 linear B cell antigen epitopes were found in the gene. At the same time, the gene was cloned into prokaryotic expression vector PGEX-4 T-1 to construct recombinant expression plasmid PGEX-4 T-1-OmpF, which was transferred into Escherichia coli BL21 to obtain OmpF recombinant protein. Western-blot analysis showed that the recombinant protein had a molecular weight of about 66 kD(contain tagged proteins) and could be recognized by Salmonella chicken positive serum. It was preliminarily confirmed that the recombinant protein had immunogenicity and laid a theoretical foundation for further research and development of Salmonella enteritidis subunit vaccine.