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细胞色素P450 3A4及谷胱甘肽S转移酶A1重组质粒的构建及表达

常彬霞 游绍莉 李保森 貌盼勇 辛绍杰

引用本文:
Citation:

细胞色素P450 3A4及谷胱甘肽S转移酶A1重组质粒的构建及表达

基金项目: 

军队临床高新技术重大项目(2010gxj097); 

详细信息
  • 中图分类号: R96

Construction and evaluation of recombinant plasmids expressing CYP3A4 and GSTA1

Research funding: 

 

  • 摘要:

    目的构建重组质粒pBudCE4.1-细胞色素P450 3A4(CYP3A4)-谷胱甘肽S转移酶A1(GSTA1),使CYP3A4和GSTA1可在真核细胞C3A中表达,并且增加其表达量。方法从含有GSTA1和CYP3A4的开放阅读框(ORF)克隆中扩增GSTA1和CYP3A4基因;将片段GSTA1以及载体pBuDCE4.1双酶切后连接并转化,质粒命名为pBuDCE4.1-GSTA1;将片段CYP3A4以及质粒pBuDCE4.1-GSTA1双酶切后连接并转化,所得重组质粒命名为pBudCE4.1-CYP3A4-GSTA1;测序验证重组克隆中目的基因片段的序列信息;构建稳定细胞系,测定CYP3A4活性及GSTA1的表达情况。结果酶切及测序验证双表达重组质粒pBudCE4.1-CYP3A4-GSTA1构建成功,CYP3A4及GSTA1在转染细胞系中的表达量增多。结论构建成的双表达重组质粒pBudCE4.1-CYP3A4-GSTA1符合应用要求。

     

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  • 出版日期:  2012-09-20
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