鼠伤寒沙门菌鞭毛蛋白与新城疫病毒F蛋白的融合表达及免疫原性分析

Coexpression of flic Protein of Salmonella typhimurium and F Protein of NDV and Its Immunogenicity Analysis in Mice

应用PCR技术分别扩增鼠伤寒沙门菌鞭毛蛋白fliC基因以及含有新城疫病毒F蛋白部分表位的基因片段,通过柔性肽(Gly4Ser)2编码序列将二者串联并克隆到质粒pET30a+上,获得重组原核表达质粒pET-fliC-F,将其转化入大肠埃希菌BL21(DE3)中表达。Western blot证实重组菌表达的fliC-F融合蛋白能与小鼠抗fliC和抗F蛋白两种多抗血清发生特异性反应。动物试验显示,fliC-F融合蛋白能够刺激C3H/HeJ小鼠产生针对F蛋白的特异性血清抗体,说明原核表达系统表达的fliC-F融合蛋白具有较好的免疫原性。

The fliC gene of Salmonella typhimurium, and the partial fragment of fusion protein （F） gene of Newcastle disease virus were amplified using PCR and ligated with a DNA sequence encoding a fh：xibte peptide （Gly4 Ser）2. The fusion gene was inserted into plasmid pET30a＋ to construct the recombinant prokaryotic expression plasmid pET-fliC-F. Then the recombinant plasmid was transformed into E. coli BL21（DE3） and induced to express the desired fliC-F protein. Western blot analysis showed that the fliC -F protein could react with murinal antiserum against fliC or F protein,respectively. It indicated that the fusion protein could significantly elicit specific immune response in immunized C3H/HeJ mice. These results demonstrated that the fliC F fusion protein expressed by E. coli had good immunogenicity.