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:
Reprint 's Address:
Email:
Source :
Year: 2006
Language: Chinese
Cited Count:
SCOPUS Cited Count:
ESI Highly Cited Papers on the List: 0 Unfold All
WanFang Cited Count:
Chinese Cited Count:
30 Days PV: 2
Affiliated Colleges: