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本文引用的文献

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Coding-sequence determinants of gene expression in Escherichia coli.大肠杆菌中基因表达的编码序列决定因素。
Science. 2009 Apr 10;324(5924):255-8. doi: 10.1126/science.1170160.
2
Expression and purification of orphan cytochrome P450 4X1 and oxidation of anandamide.孤儿细胞色素P450 4X1的表达与纯化以及花生四烯酸乙醇胺的氧化
FEBS J. 2008 Jul;275(14):3706-17. doi: 10.1111/j.1742-4658.2008.06518.x.
3
mRNA distribution and heterologous expression of orphan cytochrome P450 20A1.孤儿细胞色素P450 20A1的mRNA分布及异源表达
Drug Metab Dispos. 2008 Sep;36(9):1930-7. doi: 10.1124/dmd.108.022020. Epub 2008 Jun 9.
4
Functional expression of human cytochrome P450 enzymes in Escherichia coli.人细胞色素P450酶在大肠杆菌中的功能表达。
Curr Drug Metab. 2006 May;7(4):411-29. doi: 10.2174/138920006776873472.
5
Recombinant enzymes overexpressed in bacteria show broad catalytic specificity of human cytochrome P450 2W1 and limited activity of human cytochrome P450 2S1.在细菌中过表达的重组酶表现出人类细胞色素P450 2W1广泛的催化特异性以及人类细胞色素P450 2S1有限的活性。
Mol Pharmacol. 2006 Jun;69(6):2007-14. doi: 10.1124/mol.106.023648. Epub 2006 Mar 21.
6
Heterologous expression, purification, and properties of human cytochrome P450 27C1.人细胞色素P450 27C1的异源表达、纯化及性质
Arch Biochem Biophys. 2006 Jan 1;445(1):138-46. doi: 10.1016/j.abb.2005.11.002. Epub 2005 Nov 28.
7
Expansion of substrate specificity of cytochrome P450 2A6 by random and site-directed mutagenesis.通过随机诱变和定点诱变扩展细胞色素P450 2A6的底物特异性
J Biol Chem. 2005 Dec 9;280(49):41090-100. doi: 10.1074/jbc.M508182200. Epub 2005 Oct 7.
8
A codon window in mRNA downstream of the initiation codon where NGG codons give strongly reduced gene expression in Escherichia coli.起始密码子下游mRNA中的一个密码子窗口，其中NGG密码子在大肠杆菌中会使基因表达大幅降低。
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9
Codon bias and heterologous protein expression.密码子偏好性与异源蛋白表达
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通过密码子优化和伴侣共表达增强几种哺乳动物细胞色素P450的细菌表达。

Enhanced bacterial expression of several mammalian cytochrome P450s by codon optimization and chaperone coexpression.

作者信息

Wu Zhong-Liu, Qiao Jing, Zhang Zhi-Gang, Guengerich F Peter, Liu Yan, Pei Xiao-Qiong

机构信息

Chengdu Institute of Biology, Chinese Academy of Sciences, P.O. Box 416, 610041, Chengdu, China.

出版信息

Biotechnol Lett. 2009 Oct;31(10):1589-93. doi: 10.1007/s10529-009-0059-5. Epub 2009 Jun 26.

DOI:10.1007/s10529-009-0059-5
PMID:19557307
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4188451/
Abstract

To elucidate the effects of codon optimization and chaperone coexpression on the heterologous expression of mammalian cytochrome P450s (P450) in Escherichia coli, the expression of P450s 2B1, 2S1, 2U1, 2W1, and 27C1 were investigated. With codon optimization for N-terminus or the entire gene, the expression levels of P450 27C1, 2U1 and 2W1 increased 22-fold, 3.6-fold and 2.1-fold, respectively, while those for P450s 2B1 and 2S1 remained unchanged. With coexpression of E. coli molecular chaperones GroEL/ES, the expression level increased up to 14-fold for P450 27C1, and 3- to 5-fold for P450s 2B1, 2S1, and 2W1. Simultaneous application of these two techniques resulted in synergetic effects.

摘要

为阐明密码子优化和伴侣共表达对哺乳动物细胞色素P450（P450）在大肠杆菌中异源表达的影响，研究了P450 2B1、2S1、2U1、2W1和27C1的表达情况。对N端或整个基因进行密码子优化后，P450 27C1、2U1和2W1的表达水平分别提高了22倍、3.6倍和2.1倍，而P450 2B1和2S1的表达水平保持不变。与大肠杆菌分子伴侣GroEL/ES共表达时，P450 27C1的表达水平提高了14倍，P450 2B1、2S1和2W1的表达水平提高了3至5倍。同时应用这两种技术产生了协同效应。