摘要
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.
出处
《动物医学进展》
CSCD
北大核心
2010年第2期20-25,共6页
Progress In Veterinary Medicine
基金
国家自然科学基金资助项目(30871860)
高等学校科技创新工程重大项目(706032)
江苏省自然科学基金项目(BK2007511
BK2008011)
江苏省高校自然科学基础研究项目(07KJB230136)
江苏省高校"青蓝工程"项目
关键词
鼠伤寒沙门菌
鞭毛蛋白
新城疫病毒
融合蛋白
免疫原性
Salmonella typhimurium
fliC protein
Newcastle disclose virus fusion protein
immunogenicity