中国人兽共患病学报2011，Vol.27Issue(4)：279-282,4.
狂犬病病毒G基因的克隆表达及间接ELISA检测方法的初步建立与应用
Establishment of indirect ELISA and its application based on expressed G protein of rabies virus
摘要
Abstract
To establish indirect ELISA and detect canine sera, prokaryotical glycoprotein was expressed from rabies virus in this study. In what follows, the complete length of G gene from rabies virus was amplified by PCR using a pair of specific primers designed according to the relevant sequences from GenBank, and the PCR product was cloned into vector pGM-T.The recombinant vector was digested with BamH I and Xho Ⅰ and inserted into the prokaryotic expression vector pGEX-6P-1 to obtain the recombinant expressed vector pGEX-G and it was transferred into the E.coli strain BL21 (DE3) cells. The expression of proteins was induced by IPTG and these proteins were purified and analyzed by SDS-PAGE. And they were identified by Western blotting and the size of insert fragments was 79kDA, which could be specifically reacted with RV antiserum. The indirect ELISA assay using the purified glycoprotein as the antigen was applied in detecting 93 canine serum samples. The sensitivity of the EL ISA was 1:1 600. Comparing with the commercial ELISA kit, the accuracy of the ELISA was 96.8％.关键词
狂犬病病毒/G蛋白/表达/间接ELISA分类
医药卫生引用本文复制引用
王净,李刚,史利军,邱文英,曾妮,穆秀明,高志花,王慧文..狂犬病病毒G基因的克隆表达及间接ELISA检测方法的初步建立与应用[J].中国人兽共患病学报,2011,27(4):279-282,4.基金项目
中央级公益性科研院所基本科研业务费专项资金(2009 QN-3,2010 JS-2,0032007008),北京市科委基金项目(Z07010501780701),国家农业行业公益项目(2008-3)联合资助 （2009 QN-3,2010 JS-2,0032007008）
