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Cloning, expression and purification of isoflavone-2 '-hydroxylase from Astragalus membranaceus Bge. Var. mongolicus (Bge.) Hsiao 期刊论文

编号：

8271b727-3b5f-4eef-b733-7f53be46702d
作者：

Chen, Jing^[1] Yuan, Hui^[1] Zhang, Lin^[2] Pan, Haiyun^[3] Xu, Rongyan^[1] Zhong, Yang^[1,4] Chen, Jiakuan^[1] Nan, Peng^[1]
语种：

English
期刊：

PROTEIN EXPRESSION AND PURIFICATION ISSN：1046-5928 2015 年 107 卷 (83 - 89) ; MAR
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关键词：

Cytochrome P450 Isoflavone-2 '-hydroxylase Expression vector LC-ESI-MS/MS CO difference spectrum
摘要：

Plant cytochrome P450 enzymes play vital roles in the biosynthesis of plant secondary metabolites, including phenylpropanoids and phytoalexins. Isoflavone-2'-hydroxylase (AmI2'H) from Astragalus membranaceus Bge. Var. mongolicus (Bge.) Hsiao is a membrane protein and an eulcaryotic cytochrome P450 enzyme involved in isoflavonoid biosynthesis. We cloned the AmI2'H gene by employing RACE methods and modified the gene sequence to facilitate protein expression and increase protein solubility. Two vectors, pET-28a(+) and pCW ori(+), were used to express AmI2'H in Escherichia coli. The expression efficiency and purity of target protein were analyzed and demonstrated that pET-28a(+) vector containing the AmI2'H gene could produce larger amounts of target proteins with higher purity than pCWori(+). The purified proteins were identified as AmI2'H by LC-ESI-MS/MS analysis and their proper folding was assessed by CO difference spectrum. (C) 2014 Elsevier Inc. All rights reserved.
推荐引用方式
GB/T 7714：

Chen Jing,Yuan Hui,Zhang Lin, et al. Cloning, expression and purification of isoflavone-2 '-hydroxylase from Astragalus membranaceus Bge. Var. mongolicus (Bge.) Hsiao [J].PROTEIN EXPRESSION AND PURIFICATION,2015,107:83-89.
APA：

Chen Jing,Yuan Hui,Zhang Lin,Pan Haiyun,&Nan Peng.(2015).Cloning, expression and purification of isoflavone-2 '-hydroxylase from Astragalus membranaceus Bge. Var. mongolicus (Bge.) Hsiao .PROTEIN EXPRESSION AND PURIFICATION,107:83-89.
MLA：

Chen Jing, et al. "Cloning, expression and purification of isoflavone-2 '-hydroxylase from Astragalus membranaceus Bge. Var. mongolicus (Bge.) Hsiao" .PROTEIN EXPRESSION AND PURIFICATION 107(2015):83-89.

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