口蹄疫病毒结构蛋白VP2-3-1基因的克隆及原核表达  

cDNA Cloning of FMDV Structural Protein VP2-3-1 Gene and its Prokaryotic Expression

作　　者：李平花[1] 刘在新[1] 伏小平[2] 吴润[2] 郭建宏[1] 谢庆阁[1] 

机构地区：[1]中国农业科学院兰州兽医研究所农业部病毒学重点实验室,甘肃兰州730046 [2]甘肃农业大学,甘肃兰州730070

出　　处：《中国兽医学报》2006年第3期232-234,共3页Chinese Journal of Veterinary Science

基　　金：国家重大基础研究发展规划(973)项目(G1999011903)

摘　　要：根据口蹄疫病毒(FMDV)China99流行毒株基因序列设计特异引物,以阳性质粒pGEM-p1为模板,扩增p1基因。p1片段经Nco和Xho酶消化后,得到目的基因VP2-3-1,将其与经相同酶消化的表达载体pProEx-HTb连接并转化BL21(DE3),经PCR、酶切鉴定和序列分析筛选阳性克隆,经IPTG诱导后SDS-PAGE检测,结果表明VP2-3-1基因得到表达;Westernblotting结果显示,该表达蛋白能被口蹄疫病毒阳性血清所识别。FMDV P1 gene was amplified from positive plasmid pGEM-pl. PCR product of P1 was digested with Nco 1 and Xho 1 , and got objective gene VP2-3-1. This gene was ligated into vector pProEX-HTb and digested with Nco 1 and Xho 1 respectively, then transformed into E. coil BL21 （DE3） cells. The recombinant plasmid were identified by restriction analysis and PCR and DNA sequencing. A recombinant plasmid with FMDV VP2-3-1 gene was selected and named as pProEX-VP2-3- 1. The bacteria containing pProEX-VP2-3-1 was induced with Isopropyl-D-galactoside（IPTG） and the bacteria culture fluid were sampled and examined by SDS-PAGE and Western blotting after being properly treated. The results showed that the VP2-3-1 gene was successfully expressed in E. coil and could be recognized by the positive bovine serum to FMDV.

关 键 词：FMDV VP2—3—1基因 克隆 表达 口蹄疫 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]