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鸡细胞色素P4501A5的原核表达与活性重组

An Junfeng,Wang Yunhao,Hou Xin

Biotechnology Bulletin（2012）

Cited 0|Views35
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Abstract
旨在对鸡细胞色素P450 1A5 (CYP1A5)蛋白进行体外功能研究,采用大肠杆菌系统进行CYP1A5的异源表达.以鸡的cDNA为模板,扩增出CYP1A5基因,将该基因的N端编码区进行修饰,并连接到pCW载体中构建His-CYP1A5,经IPTG诱导在大肠杆菌中表达.经CO-差示光谱检测,所获得的His-CYP1A5具有典型的P450吸收峰.该蛋白与细胞色素P450还原酶(CPR)进行体外重组,构成的重组酶系表现出乙氧基试卤灵-O-脱乙基酶活性.结果表明,所采用的表达策略可以成功产生出具有催化活性的鸡细胞色素P4501A5( CYP1A5)蛋白.
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Cytochrome P450 1A5(CYP1A5) Heterologous expression Reconstitution in vitro Enzymatic activity
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