牛病毒性腹泻病毒E2基因生物信息学分析及可溶性表达  

BIOINFORMATICS ANALYSIS AND PROKARYOTIC EXPRESSION OF BOVINE VIRAL DIARRHEA VIRUS E2 GENE

作　　者：赵洪哲 李新培[2] 范峻豪 宋前进 张静[1] 关平原[1] 温永俊[1,2] ZHAO Hong-zhe;LI Xin-pei;FAN Jun-hao;SONG Qianjin;ZHANG Jing;GUAN Ping-yuan;WEN Yongjun(Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Diseases,Ministry of Agriculture and Rural Affairs,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Institute of Special Products,CAAS,Changchun 130112,China)

机构地区：[1]内蒙古农业大学兽医学院农业农村部动物疾病临床诊疗技术重点实验室,呼和浩特010018 [2]中国农业科学院特产研究所,长春130112

出　　处：《中国动物传染病学报》2020年第1期63-67,共5页Chinese Journal of Animal Infectious Diseases

基　　金：内蒙古农业大学高层次人才引进项目(NDGCC2016-22)

摘　　要：为了获得牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)E2基因的最佳可溶性蛋白,本试验根据GenBank已公布的BVDV E2基因序列,利用DNAStar等软件进行生物信息学分析,截取抗原性和亲水性较高的序列进行稀有密码子优化,串联信号肽序列后合成并连接到原核表达载体pET-32a(+),构建重组质粒pET-32a-Opti-E2;转化至E.coli BL21(DE3)宿主菌,经SDS-PAGE和Western blot分析,成功获得大小为43 kDa的BVDV E2基因的最佳可溶性蛋白,且该蛋白特异性较好。本试验E2蛋白的成功表达可为后续ELISA方法的建立和亚单位疫苗的开发奠定基础。The E2 gene sequence of Bovine viral diarrhea virus(BVDV)published in GenBank was analyzed using bioinformatics software such as DNA star.The high antigenic and hydrophilic sequences were intercepted and optimized with rare codons for design of tandem signal peptide sequences and synthesis.The recombinant plasmid pET-32 a-Opti-E2 was constructed and transformed into E.coli BL21(DE3)for prokaryotic expression with IPTG induction.The obtained recombinant E2 protein was purified using nickel columns and confirmed as examined in SDS-PAGE and Western blotting.The soluble BVDV E2 protein of molecular weight at 43 kDa was visualized in Western blot.The availability of the recombinant BVDV E2 protein laid the foundation for development of an ELISA method and a subunit vaccine.

关 键 词：牛病毒性腹泻病毒 囊膜结构蛋白E2 原核表达 

分 类 号：S852.659.6[农业科学—基础兽医学]