瑟氏泰勒虫p33基因与牛IL-18基因的串联表达  

Prokaryotic expression of IL-18 gene of bovine and p33 gene of Theileria sergenti

作　　者：海旭南[1] 宋建臣[1] 李海峰[1] 薛书江[1] 许应天[1] 

机构地区：[1]延边大学农学院,吉林延吉133002

出　　处：《中国兽医学报》2016年第9期1527-1530,共4页Chinese Journal of Veterinary Science

基　　金：吉林省教育厅"十二五"科学技术研究资助项目(2015-16)

摘　　要：为研究牛IL-18基因对瑟氏泰勒虫p33疫苗的免疫佐剂效应,根据GenBank上已经发表的p33基因序列和牛IL-18基因序列(IL-18)设计2对特异性引物,以瑟氏泰勒虫基因组DNA和pMD-19-T-IL-18质粒DNA为模板,采用SOE-PCR技术将2个基因连接在一起,经BamHⅠ、XhoⅠ双酶切,获得1 416bp的目的基因IL-18-p33,克隆于原核表达载体pGEX-4T-1中,构建了双基因重组表达载体pGEX-4T-1-IL-18-p33,转化大肠杆菌BL21,经IPTG诱导,表达出预期大小为79 000的融合蛋白。Western blotting检测结果显示,该蛋白与瑟氏泰勒虫抗血清呈阳性反应,表明融合蛋白具有反应原性,为进一步研究IL-18对瑟氏泰勒虫p33基因疫苗的免疫佐剂效应奠定了基础。In order to study the immune adjuvant effect of IL-18 gene,two pair of primers were designed according to the p33 gene of Theileria sergenti and IL-18 gene of cattle in GenBank. Theileria sergenti p33 gene was linked with cattle IL-18 gene by SOE-PCR and then inserted into pMD19-T simple vector. The 1 416 bp fusion gene was cloned into pGEX-4T-1 plasmid by rstriction endonuclease BamH I and Xho I and transformed into E. coli BL21 for expression. SDS-PAGE and Western blotting analysis showed that the recombinant fusion protein had a molecular weight of 79 000 and could be recognized by positive serum of Theileria sergenti. The results showed that the fusion protein has a reactogenicity,and it would establish the foundation for further study on the immune adjuvant effect of IL-18 gene.

关 键 词：瑟氏泰勒虫 p33基因 牛IL-18基因 串联表达 

分 类 号：S852.7[农业科学—基础兽医学]