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Construction of expressing vector of pSUPER-shRNA/mrp1 and its expression in vitro
Author(s): PAN Guang-dong, YAN Lü-nan, YANG Jian-qing, CHU Guang-ping, LIU Qiang, XIAO Yi
Pages: 48-
53
Year: 2010
Issue:
1
Journal: Chinese Journal of Hepatobiliary Surgery
Keyword: Gene expression; Multidrug resistance; Plasmid;
Abstract: 目的 构建shRNA/mrp1的质粒表达载体并验证其体外表达效率.方法 根据业已筛选出的mrp1基因RNAi靶序列,按照pSUPER的设计要求合成64 bp的寡核苷酸序列,将其退火后形成双链并用双酶切法克隆到pSUPER得到质粒pSUPER-shRNA/mrp1,转染感受态大肠杆菌,筛选阳性克隆,经测序证实后扩增培养并以去内毒素试剂盒提取,然后转染HepG2/mrp1细胞,阴性载体为对照组,Rea...
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