安徽农业科学2017，Vol.45Issue(12)：122-123,129,3.
牛病毒性腹泻黏膜病毒E2基因的原核表达与免疫原性分析
Prokaryotic Expression and Inmunogenicity of E2 Gene of BVDV
摘要
Abstract
[Objective]To prokaryotic express the bovine viral diarrhea-muscosal disease viruses E2 gene encoding protein.[Method]Bovine viral diarrhea-muscosal disease viruses E2 gene were amplified by PCR and linked into the pET-32a prokaryotic expression vector,prokaryotic expression recombinant plasmid pET32a-E2 was constructed,which was then trandformed into E.coli(Rosetta)cells for protein expression.E2 protein of recombinant strains was induced to express by 1 mmol/L IPTG and SDS-PAGE electrophoresis,target protein was purified by Ni-NTA affinity chromatography column and the immunogenicity was identified by Western blot analysis.[Result]The recombinant plasmid pET32a-E2 was confirmed by PCR,restriction enzyme.It had high-level expression in E.coli.SDS-PAGE showed that recombinant protein with molecular weight of 58 kDa,with concerntration of 0.521 mg/mL.Western blot showed that recombinant protien can reacts with positive serum,indicating good immunogenicity.[Conclusion]E2 protein is expressed in successfully,which lays foundation for establishing BVDV detection method in future.关键词
牛病毒性腹泻黏膜病毒/E2基因/原核表达Key words
Bovine viral diarrhea-mucosal disease viruses/E2 gene/Prokaryotic expression分类
农业科技引用本文复制引用
王宇婷,马莉莉,李晓月,王士霞,毕莹,倪宏波..牛病毒性腹泻黏膜病毒E2基因的原核表达与免疫原性分析[J].安徽农业科学,2017,45(12):122-123,129,3.基金项目
黑龙江省农垦总局"十二五"重点科技攻关项目(HNK125B-11-10A、HNK125B-11-02). （HNK125B-11-10A、HNK125B-11-02）
