动物医学进展Issue(12)：17-21,5.
猪繁殖与呼吸综合征病毒 GP5基因的克隆和原核表达
Cloning and Prokaryotic Expression of PRRSV GP5 Gene
摘要
Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV)can cause obstacle of all age pigs in respiratory system and dysfunction of sow reproduction.The GP5 of PRRSV is important to virus adhe-sion,pathogenicity,immune protection.In this study,we extracted the RNA of PRRSV genome.The full-length GP5 gene was amplified from cDNA of PRRSV by reverse transcription PCR.The GP5 gene fragment is 594 bp.The GP5 gene was subcloned into the prokaryotic expression vector pGEX-6P-1.By colony PCR identification and restriction endonuclease digestion,the recombinant plasmid (pGEX-6P-1-GP5)was transformed into E.coli BL21(DE3),His-GP5 protein was expressed by isopropy-beta-D-thio-galactopyranoside (IPTG )induction,whose molecular weight is 41 ku.The results showed that His-GP5 protein was mainly in the form of inclusion bodies and more than 35% of the total bacterial proteins by SDS-PAGE analysis.The purification products of recombinant protein was more than 75% of the total protein.The recombinant protein has preferably an immunogenic by Western-blot analysis.This research is the foundation to the preparation of the anti-GP5 antibody and genetically engineered vaccine of GP5.关键词
猪繁殖与呼吸综合征病毒/GP5 基因/原核表达/纯化Key words
PRRSV/GP5 gene/prokaryotic expression/purification分类
农业科技引用本文复制引用
袁庄川,王鲁彦,梅匀安,张敏,王晓杜..猪繁殖与呼吸综合征病毒 GP5基因的克隆和原核表达[J].动物医学进展,2013,(12):17-21,5.基金项目
中国博士后基金项目（2012M521187）；浙江农林大学人才启动基金 （）
