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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。

     

  • [1]Pachecka J, Tomaszewski P, Kubiak-Tomaszewska G.Cy-tochrome P450 polymorphism-molecular, metabolic andpharmacogenetic aspects I.Mechanisms of activity of cyto-chrome P450 monooxygenases[J].Acta Pol Pharm, 2008, 65 (3) :303-306.
    [2] Oakley A. Glutathione transferases: a structural perspective[J]. Drug Metab Rev, 2011, 43 (2) : 138-151.
    [3]Bertz RJ, Granneman GR.Use of in vitro and in vivo datatoestimate the likeli-hood of metabolic pharmacokinetic inter-actions[J].Clin Pharmacokinet, 1997, 32 (3) :210-258.
    [4]Tomaszewski P, Kubiak-Tomaszewska G, Pachecka J.Cy-tochrome P450 polymorphism-molecular, metabolic, andpharmacogenetic aspects II.Participation of CYP isoenzymesin the metabolism of endogenous substances and drugs[J].Acta Pol Pharm, 2008, 65 (3) :307-318.
    [5] Gertz M, Harrison A, Houston JB, et al. Prediction of human intestinal first-pass metabolism of 25 CYP3A substrates from in vitro clearance and permeability data[J]. Drug Metab Dispos, 2010, 38 (7) : 1147-1158.
    [6]Rowe JD, Nieves E, Listowsky I.Subunit diversity and tis-sue distribution of human glutathione S-transferases:inter-pretations based on electrospray ionization-MS and peptidesequence-specific antisera[J].Biochem J, 1997, 325 (pt2) :481-486.
    [7] Whalen R, Boyer T. Human glutathione-S-transferase[J]. Semin Liver Dis, 1998, 18 (4) : 345-358.
    [8]Juronen E, Tasa M, Uuskula M, et al.Purification, charac-terization and tissue distribution of human class theta glutathi-one S-transferaseT1-1[J].Bilchem Mol Biol Int, 1996, 39 (1) :21-29.
    [9] Farrell G. Effects of disease on expression and regulation of CYPs[J]. Mol Aspects Med, 1999, 20 (1-2) : 55-70.
    [10]Yang LQ, Li SJ, Cao YF, et al.Different alterations of cyto-chrome P450 3A4 isoform and its gene expression in livers ofpatients with chronic liver diseases[J].World J Gastroen-terol, 2003, 9 (2) :359-363.
    [11]Chalasani N, Gorski JC, Patel NH, et al.Hepatic and intesti-nal cytochrome P450 3A activity in cirrhosis:effects of tran-sjugular intrahepatic portosystemic shunts[J].Hepatology, 2001, 34 (6) :1103-1108.
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  • 出版日期:  2012-09-20
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