口蹄疫病毒非结构蛋白3ABC基因的原核表达及其产物的活性检测  

Prokaryotic expression of FMDV non-structural protein 3ABC gene and activities of the expressed product

作　　者：曲哲会[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2006年第9期696-700,共5页Chinese Veterinary Science

摘　　要：利用分别带有BamHⅠ和HindⅢ酶切位点的引物扩增出FMDV 3ABC基因,将其克隆到pMD18-T载体。经BamHⅠ和SalⅠ酶消化后,克隆到表达载体pGEX-6p-1上,构建重组质粒pGEX-3ABC。将该重组质粒转化到大肠埃希氏菌BL21(DE3)pLysS,经终浓度1 mmol/L的IPTG诱导,获得约70 ku的融合蛋白GST-3ABC。Western-blotting结果显示,表达的融合蛋白GST-3ABC可以与感染FMDV的牛血清发生免疫反应。以纯化的GST-3ABC蛋白为抗原的间接ELISA检测结果与以VP2蛋白为抗原的间接ELISA检测结果相比较,表明,融合蛋白GST-3ABC可以用于鉴别感染FMDV和疫苗免疫的牛血清。The 3ABC gene of FMDV was amplified by primers with digestion sites of restriction endonuclease BarnH Ⅰ and Hind Ⅲ, respectively. The amplified fragment was cloned into pMD18-T vector. The recombinant plasmid pT-3ABC was digested by BarnH Ⅰ and Sal Ⅰ and then cloned into pGEX-6p-1. The recombinant plasmid pGEX-3ABC was transformed into BL21 （DE3） plysS and the target protein was induced by IPTG. The expressed fusion protein was identified by SDS-PAGE and Western blotting. Results showed that the expressed protein could react with FMDV-infected bovine serum. Compared to VP2- ELISA, the 3ABC-ELISA results could differentiate the infected animals from the animals immunized with the vaccine against FMDV.

关 键 词：口蹄疫病毒 3ABC基因 原核表达 鉴别诊断 

分 类 号：S852.659.6[农业科学—基础兽医学]