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表达外源绿荧光蛋白重组山羊痘病毒的构建及纯化
  • 摘要

    以我国自行培养并得到广泛应用的山羊痘病毒(Goatpox virus,GPV)疫苗株为亲本,以复制非必需胸苷激酶(tk)基因作为外源基因插入靶点及同源重组臂.利用转移载体pTK-lacZ-gfp及其与GPV发生同源重组获得的病毒(rGPV-lacZ-gfp)验证外源基因β-半乳糖苷酶(lacZ)和绿荧光蛋白(gfp)基因在背靠背启动子p11和p7.5启动下成功稳定表达后,将转移载体lacZ基因更换为gpt(黄嘌呤鸟嘌呤磷酸转移酶)基因(pTK-gpt-gfp),利用MPA阻断核酸代谢途径筛选并获得了遗传稳定表达的单一纯化病毒克隆(rGPV-gpt-gfp).研究建立了表达外源基因重组GPV构建系统及其克隆纯化方法,为进一步开展GPV活载体疫苗研制及相关基础研究提供了技术平台.

  • 作者

    程小文  胡森  王喜军  鲍恩东  步志高  CHENG Xiao-wen  HU Sen  WANG Xi-jun  Bao En-Dong  BU Zhi-gao 

  • 作者单位

    中国农科院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨,150001;南京农业大学动物医学院,南京,210095/中国农科院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨,150001/南京农业大学动物医学院,南京,210095

  • 刊期

    2006年5期 ISTIC PKU

  • 关键词

    重组山羊痘病毒  β-半乳糖苷酶  绿荧光蛋白  黄嘌呤鸟嘌呤磷酸转移酶 

参考文献
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