Development of an anhydrotetracycline-inducible gene expression system for solvent-producing Clostridium acetobutylicum: A useful tool for strain engineering

被引：46

作者：

Dong, Hongjun ^{[1
]}

Tao, Wenwen ^{[1
]}

Zhang, Yanping ^{[1
]}

Li, Yin ^{[1
]}

机构：

[1] Chinese Acad Sci, Inst Microbiol, Beijing 100101, Peoples R China

来源：

METABOLIC ENGINEERING | 2012年 / 14卷 / 01期

基金：

国家高技术研究发展计划(863计划);

关键词：

Clostridium acetobutylicum; Inducible gene expression systems; Tetracycline regulatory system; Metabolic engineering tools; COUNTER-SELECTABLE MARKER; ESCHERICHIA-COLI; TET REPRESSOR; PROTEIN; INACTIVATION; MAZF; CONSTRUCTION; MANIPULATION; PROMOTERS; THIOLASE;

D O I：

10.1016/j.ymben.2011.10.004

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Clostridium acetobutylicum is an important solvent (acetone-butanol-ethanol) producing bacterium. However, a stringent, effective, and convenient-to-use inducible gene expression system that can be used for regulating the gene expression strength in C acetobutylicum is currently not available. Here, we report an anhydrotetracycline-inducible gene expression system for solvent-producing bacterium C acetobutylicum. This system consists of a functional chloramphenicol acetyltransferase gene promoter containing tet operators (tetO), Pthl promoter (thiolase gene promoter from C acetobutylicum) controlling TetR repressor expression cassette, and the chemical inducer anhydrotetracycline (aTc). The optimized system, designated as pGusA2-2tetO1, allows gene regulation in an inducer aTc concentration-dependent way, with an inducibility of over two orders of magnitude. The stringency of TetR repression supports the introduction of the genes encoding counterselective marker into C acetobutylicum, which can be used to increase the mutant screening efficiency. This aTc-inducible gene expression system will thus increase the genetic manipulation capability for engineering C acetobutylicum. (C) 2011 Elsevier Inc. All rights reserved.

引用

页码：59 / 67

页数：9

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