摘要
将口蹄疫病毒(FMDV)的VP1基因,通过pPROex-HT表达载体在大肠杆菌BL21(DE3)中成功表达,获得大小为31ku的融合蛋白,Westernblot检测证实表达的该蛋白具有良好的生物学活性。以纯化的融合蛋白为抗原建立了猪FMDVVP1蛋白间接ELISA检测方法。通过对80份田间血清样品的检测表明,该方法与FMDV液相阻断ELISA(国标试剂盒)的总符合率为96.25%,表明建立的VP1蛋白间接ELISA检测方法具有很好的特异性和敏感性。
The complete gene encoding the structural protein of FMDV（VP1）was subcloned into expression vector pPROex-HT,resulting in the fusion expression plasmid pPROexHT-VP1.After transformed into E.coli BL21（DE3）and induced by IPTG,the fusion protein was expressed in high level.Western blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV.Based on the fusion protein further purified,a novel indirect ELISA（VP1-ELISA）was developed to detect FMDV antibody in pigs.Comparison between VP1-ELISA and the government standard kit（liquid phase block ELISA）showed the two methods had 96.25 percent agreement by detecting 80 serum samples,indicating that the indirect VP1-ELISA was specific and sensitive.
出处
《生物工程学报》
CAS
CSCD
北大核心
2007年第5期961-966,共6页
Chinese Journal of Biotechnology
基金
国家重点基础研究发展计划(No.2005CB523201)
国家支撑计划(2006BAD06A03)~~
关键词
口蹄疫病毒抗体
VP1蛋白
间接ELISA
antibodies against foot-and-mouth disease virus,VP1 protein,indirect ELISA
