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鸡MHC Iα-生物素化序列融合基因的构建、表达及纯化
  • 摘要

    实验克隆了莱航鸡(Gallusgallus)MHC Iα(BF2)(GenBank登陆号:AY989897)全基因,分析了其信号肽序列,构建了缺失信号肽且拼接了BirA底物肽序列(BSP)的融合蛋白重组表达载体pET-BF2-BSP,在大肠杆菌(Escherichia coli)中得到表达,并优化了诱导剂浓度、起始诱导菌体浓度及诱导时间等表达条件.SDS-PAGE分析结果表明所得融合目的蛋白约为40 kD,Western-blot结果显示该重组蛋白成功融合了6×His标签;pET-BF2-BSP最优化表达条件分别为:菌体起始诱导浓度OD600nm0.8~1.2,IPTG诱导浓度1.1 mmol/L,诱导时间2~4 h;建立了该重组蛋白变性状态下通过镍柱亲和层析纯化包涵体的方法.实验获得了高纯度的在体外构建鸡MHC-肽四聚体所需的重组蛋白BF2-BSP.

  • 作者

    刘光亮  童铁钢  王群  肖一红  孟庆文  吴东来  LIU Guang-liang  TONG Tie-gang  WANG Qun  XIAO Yi-hong  MENG Qing-wen  WU Dong-lai 

  • 作者单位

    中国农业科学院哈尔滨兽医研究所,哈尔滨,150001;中国农业科学院研究生院,北京,100081/中国农业科学院哈尔滨兽医研究所,哈尔滨,150001

  • 刊期

    2006年5期 ISTIC PKU

  • 关键词

    鸡MHC克隆  生物素化序列  表达  纯化 

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