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author:

赖庆安 (赖庆安.) [1] | 刘树滔 (刘树滔.) [2] | 卢菀华 (卢菀华.) [3] | 陈莉 (陈莉.) [4] | Toshihide NISHIMURA (Toshihide NISHIMURA.) [5] | 饶平凡 (饶平凡.) [6]

Abstract:

氨肽酶H是生物组织内一种重要的蛋白质端解酶。本文从鸡肝组织中提取总RNA反转录成 cDNA第一链,根据APH基因序列设计合成特异性引物,PCR扩增APH基因,并插入原核表达载体 pGEM(-T Easy,转化感受态受体菌XL1-Blue,以蓝白菌落筛选转化株,用PCR检测白色菌落,确认获得了含cDNA的阳性重组子。之后,该DNA再亚克隆到载体pET22b(+)的NdeⅠ和BamHⅠ位点上。然后转化大肠杆菌Rosetta(DE3),筛选阳性菌株,酶切和测序结果显示APH基因已正确插入,用 IPTG(异丙基-β-D-硫代半乳糖苷)进行APH的诱导表达,在SDS-PAGE的52kD处有一条明显的新增...

Keyword:

基因克隆 氨肽酶H(APH) 纯化 表达

Community:

  • [ 1 ] 福州大学生物科学与工程学院生物工程研究所
  • [ 2 ] Graduate School of Biosphere Science,Hiroshima University,Japan

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Year: 2006

Language: Chinese

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ESI Highly Cited Papers on the List: 0 Unfold All

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30 Days PV: 3

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