中国兽医科学2018，Vol.48Issue(6)：671-677,7.DOI:10.16656/j.issn.1673-4696.2018.0123
蓝舌病毒VP5蛋白抗体的制备及鉴定
Preparation and identification of antibodies against VP5 protein of bluetongue virus
摘要
Abstract
In this study, the S6 full-length gene coding for the BTV-16 VP5 protein was synthesized based on the sequence of the bluetongue virus type 16(BTV-16) standard strain RSAvvvv in GenBank, and then the synthesized sequence was ligated with the pUC-57 vector. Apair of primers was designed to amplify the VP5Δ41aa gene with the deletion of N-terminal 1-41 amino acids by PCR, and the VP5Δ41aa gene was subcloned into pET-32a(+) vector to construct the recombinant expression plasmid pET-VP5Δ41aa. The recombinant plasmid was then transformed into E. coli BL21(DE3)and induced with IPTG. SDS-PAGE results showed that the recombinant VP5Δ41aa protein(rVP5Δ41aa) was expressed and purified successfully by His-Tag purification resin, and polyclonal antibodies was preparaed by immunizing New Zealand white rabbits with rVP5Δ41aa. The Western-blot results showed that the VP5Δ41aa antibodies can react with VP5Δ41aa expressed in E. coli as well as the VP5 protein in BTV-infected cells. The antibodies can also recognize the VP5 protein with native conformation in BTV-infected C6/36 cells by immunofluorescence assay. In conclusion, this VP5Δ41aa antibodies would be useful to further study the function of VP5 protein.关键词
蓝舌病毒/VP5基因/原核表达/多克隆抗体/鉴定Key words
bluetongue virus/VP5 gene/prokaryotic expression/polyclonal antibody/identification分类
农业科技引用本文复制引用
康棣,独军政,高闪电,田占成,Darien Kheder ALI MOHAMED,郭艳妮,刘光远,罗建勋,殷宏..蓝舌病毒VP5蛋白抗体的制备及鉴定[J].中国兽医科学,2018,48(6):671-677,7.基金项目
国家重点研发计划项目(2017YFD0502304) （2017YFD0502304）
国家自然科学基金项目(31672562) （31672562）
