人细胞色素P450氧化还原酶的克隆表达和抗体的制备

doi:10.3969/j.issn.1000-1565.2006.01.018

摘要/Abstract

摘要： 细胞色素P450还原酶(CPR)是人细胞色素P450酶系的电子供给者,对后者活性的发挥起到重要作用.本研究将从人胚胎cDNA文库中得到的人CPR的编码基因,连接入pT7450表达载体中,在大肠杆菌BL21(DE3)中高效表达并纯化.每升发酵液可得到纯化蛋白49 mg,占总蛋白含量的10%,以细胞色素C为底物检测酶活性,比活为65 u/mg.利用纯化的CPR作为抗原,常规免疫纯系大耳家兔,获得高效价、高特异性的抗血清,抗体滴度为1:200 000(EUSA法),纯化后抗体应用于不同组织中CPR含量的评价,证明重组CPR及其抗体可用于细胞色素P450的体外药物代谢及细胞色素P450与CPR作用机理的研究.

关键词: 人细胞色素P450还原酶, 克隆, 表达, 抗体, 药物代谢

中图分类号:

Q786

程婕,龚毅,陈悦,杨凌. 人细胞色素P450氧化还原酶的克隆表达和抗体的制备[J]. 河北大学学报(自然科学版), 2006, 26(1): 70-76.

CHENG Jie,GONG Yi,CHEN Yue,YANG Ling. Cloning and Expression of Human Cytochrome P450 Reductase and Production of Polyclonal Antibodies Against the Recombinant Protein[J]. Journal of Hebei University (Natural Science Edition), 2006, 26(1): 70-76.

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