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猪瘟病毒贵州流行株E2基因原核表达及其免疫原性的研究
引用本文:汤德元,李春燕,罗险峰,黄涛,曾智勇,甘振磊,王凤.猪瘟病毒贵州流行株E2基因原核表达及其免疫原性的研究[J].畜牧兽医学报,2011,42(11).
作者姓名:汤德元  李春燕  罗险峰  黄涛  曾智勇  甘振磊  王凤
作者单位:1. 贵州大学动物科学学院,贵阳,550025
2. 贵州省动物疫病预防控制中心,贵阳,550008
基金项目:贵州省省长基金项目,贵州省2011年农业攻关项目
摘    要:本研究根据GenBank登录的猪瘟病毒Shimen和C株的E2基因序列设计1对特异引物,采集来自贵州的疑似猪瘟病死猪组织,提取总RNA,进行RT-PCR扩增,将扩增产物测序后进行核苷酸序列比较分析.并将RTPCR产物克隆到pMD18-T载体上,构建重组质粒pMD18-T-E2,经双酶切鉴定、核苷酸序列分析后,将E2基因亚克隆到pET-32a(+)原核表达载体上,成功构建猪瘟病毒E2基因原核表达质粒pET-32a-E2,将构建好的原核表达质粒pET-32a-E2转化至宿主菌BL21中,诱导表达目的蛋白,表达产物经SDS-PAGE电泳、Western blot分析,得到了约58 ku的条带,与预期大小相符,进一步提取、纯化目的蛋白,将目的蛋白加入弗氏佐剂免疫小鼠,免疫后采血检测抗体,结果显示目的蛋白能诱导小鼠产生一定抗体水平,该研究为猪瘟亚单位疫苗的研究奠定了基础.

关 键 词:猪瘟病毒  E2基因  原核表达  免疫原性

Prokaryotic Expression and Immunogenicity of E2 Gene of CSFV Guizhou Epidemic Strain
TANG De-yuan,LI Chun-yan,LUO Xian-feng,HUANG Tao,ZENG Zhi-yong,GAN Zhen-lei,WANG Feng.Prokaryotic Expression and Immunogenicity of E2 Gene of CSFV Guizhou Epidemic Strain[J].Acta Veterinaria et Zootechnica Sinica,2011,42(11).
Authors:TANG De-yuan  LI Chun-yan  LUO Xian-feng  HUANG Tao  ZENG Zhi-yong  GAN Zhen-lei  WANG Feng
Institution:TANG De-yuan1,LI Chun-yan1,LUO Xian-feng2,HUANG Tao1,ZENG Zhi-yong1,GAN Zhen-lei1,WANG Feng1 (1.Department of Animal Science,Guizhou University,Guiyang 550025,China,2.Animal Disease Prevention and Control Center in Guizhou Province,Guiyang 550008,China)
Abstract:According to published cDNA sequence of E2 gene in GenBank of CSFV Shimen and C strain,the synthesized specific primers was designed.E2 gene fragments of classical swine fever virus Guizhou strain were amplified by extraction tissue RNA of the suspected CSF dead and RT-PCR.The PCR products were sequenced and comparatively analyzed.The products of RT-PCR were cloned into pMD18-T vector to form recombinants pMD18-T-E2 and subcloned into prokaryotic expressing vector pET-32a(+).The pET-32a-E2 was transformed i...
Keywords:CSFV  E2 gene  prokaryotic expression  immunogenicity  
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