动物医学进展Issue(10)：60-63,64,5.
高致病性PRRSV JL-04／12株核衣壳蛋白的表达与抗原性分析
Expression and Antigenicity Analysis of Nucleocapsid Protein from Highly Pathogenic PRRSV JL-0 4/1 2 Strain
摘要
Abstract
In order to obtain high concentration and efficient prokaryotic expression protein as antigen for ELISA diagnosis,the nucleocapsid protein (N protein ) gene of porcine reproductive and respiratory syndrome virus (PRRSV)was amplificated by RT-PCR and cloned into prokaryotic expression vector pGEX-6P-1.The cloned products were transformed into Escherichiacoli BL-21 competent cells.The expression of recombinant fusion pro-tein was induced by IPTG in E.coli BL21(DE3)system.The efficient expression of recombinant proteins was ob-tained by the temperature and IPTG concentration optimization.The expressed protein was detected by SDS-PAGE and Western blot,and purified with GST column.The immunogenicity of N protein was examined in mice.The re-sults showed that the expression of recombinant protein was efficiently induced under the condition:0.1 mmol/L IPTG,28℃ and 4 h.The target protein was in the supernatant and accounted for 40% of the total protein.The prokaryotic expressed recombinant N protein can induce the production of antibody in mice,and the antibody titer was 1∶2 5 6 .Prokaryotic expression N protein had the same antigenicity with PRRSV.关键词
猪繁殖与呼吸综合征病毒/核衣壳蛋白/原核表达/纯化Key words
Porcine reproductive and respiratory syndrome virus/nucleocapsid protein/prokaryotic expression/purification分类
农业科技引用本文复制引用
王凤雪,刘莹,杨勇,朱洪伟,孙娜,刘杏,程世鹏,温永俊..高致病性PRRSV JL-04／12株核衣壳蛋白的表达与抗原性分析[J].动物医学进展,2015,(10):60-63,64,5.基金项目
吉林省科技发展计划项目（20121823）；国家“十二五”高技术研究发展计划项目 （）
