水疱性口炎病毒两血清型G蛋白的截短表达和间接ELISA方法的建立  

Establishment of an Indirect ELISA for Testing Antibodies Against the Two Types of Vesicular Stomatitis Virus Based on Truncated G Protein Expression

作　　者：臧京帅[1] 高志强[2] 张乐萃[1] 张鹤晓[2] 

机构地区：[1]青岛农业大学动物科技学院,山东青岛266109 [2]北京出入境检验检疫局,北京朝阳100026

出　　处：《中国兽医杂志》2014年第6期69-72,共4页Chinese Journal of Veterinary Medicine

基　　金：质检公益性行业科研专项"马重大外来虫媒病抗体;核酸快速检测技术研究"(201010033)

摘　　要：根据水疱性口炎病毒印第安那型(IND型)和新泽西型(NJ型)G蛋白抗原表位分布图,在表达抗原表位较密集的基因区域设计引物,采用RT-PCR方法分别扩增水泡性口炎病毒IND型657 bp G蛋白基因片段和NJ型666 bp G蛋白基因片段,并将其克隆到pET-32a表达载体。将重组质粒转化宿主菌Rosetta,经1.0 mmol/L IPTG诱导,外源基因获高效表达。通过Western-blot以及ELISA试验证明截短表达的2个血清型的重组G蛋白与标准阳性血清发生反应。将2个血清型的重组G蛋白等量混合作为检测抗原包被酶标板,建立了可检测IND型和NJ型水泡性口炎病毒抗体的间接ELISA方法。The primers at the epitope dense region of G protein （Gpr） of vesicular stomatitis virus （VSV） Indiana （IND） type and New Jersey type（N J） were designed. After analyzing the antigen epitopes of IND and NJ VSV Gpr, the truncated Gpr gene fragments of 657 bp （IND） and 666 bp （N J） were amplified by RT-PCR, and was then inserted into pET-32a expression vector. To obtain the functional truncated G protein, the recombinant plasmids were transformed into E. coli Rosetta, and induced by 1.0 mmol/L IPTG induction. The results of SDS page showed that the exogenous gene was highly expressed. The Western-blot was carried out to confirm that the shortened recombinant expression protein Gpr could react with the standard positive serum. Furthermore, the recombinant truncated G protein of two serotypes were equally mixed to coat ELISA plates, and indirect ELISA method was established for testing antibodies against VSV of IND and NJ type.

关 键 词：水疱性口炎病毒 G蛋白基因 重组表达 间接ELISA 

分 类 号：S852.65[农业科学—基础兽医学]