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Position: Home > Articles > Prokaryotic expression of IL-18 gene of bovine and p33 gene of Theileria sergenti Chinese Journal of Veterinary Science 2016 (9) 1527-1530

瑟氏泰勒虫p33基因与牛IL-18基因的串联表达

作  者:
海旭南;宋建臣;李海峰;薛书江;许应天
单  位:
延边大学农学院
关键词:
瑟氏泰勒虫;p33基因;牛IL-18基因;串联表达
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。
译  名:
Prokaryotic expression of IL-18 gene of bovine and p33 gene of Theileria sergenti
作  者:
HAI Xu-nan;SONG Jian-chen;LI Hai-feng;XUE Shu-jiang;XU Ying-tian;Agricultural College of Yanbian University;
关键词:
Theileria sergenti;;p33 gene;;interleukin-18(IL-18)gene of cattle;;fusion gene expression
摘  要:
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-1plasmid by rstriction endonuclease BamHⅠand XhoⅠand 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.

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