title: Development of an inducible transposon system for efficient random mutagenesis in Clostridium acetobutylicum.
abstract: Clostridium acetobutylicum is an industrially important Gram-positive organism, which is capable of producing economically important chemicals in the ABE (Acetone, Butanol and Ethanol) fermentation process. Renewed interests in the ABE process necessitate the availability of additional genetics tools to facilitate the derivation of a greater understanding of the underlying metabolic and regulatory control processes in operation through forward genetic strategies. In this study, a xylose inducible, mariner-based, transposon system was developed and shown to allow high-efficient random mutagenesis in the model strain ATCC 824. Of the thiamphenicol resistant colonies obtained, 91.9% were shown to be due to successful transposition of the catP-based mini-transposon element. Phenotypic screening of 200 transposon clones revealed a sporulation-defective clone with an insertion in spo0A, thereby demonstrating that this inducible transposon system can be used for forward genetic studies in C. acetobutylicum.
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Shu
authors.initials: S
authors.lastname: Xu
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Changsheng
authors.initials: C
authors.lastname: Chai
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China Jiangsu National Synergetic Innovation Center for Advanced Materials, SICAM, 200 North Zhongshan Road, Nanjing 210009, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China Jiangsu National Synergetic Innovation Center for Advanced Materials, SICAM, 200 North Zhongshan Road, Nanjing 210009, China.
authors.forename: Weihong
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Minton
authors.affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China Shanghai Collaborative Innovation Center for Biomanufacturing Technology, 130 Meilong Road, Shanghai 200237, China ygu02@sibs.ac.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Gu
mesh_terms: D000096:Acetone / Q000378:metabolism; D000440:Butanols / Q000378:metabolism; D046969:Clostridium acetobutylicum / Q000235:genetics* / Q000378:metabolism*; D004251:DNA Transposable Elements / Q000235:genetics*; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation / Q000235:genetics*; D016296:Mutagenesis / Q000235:genetics; D013171:Spores, Bacterial / Q000235:genetics*; D014994:Xylose / Q000378:metabolism
chemical_list: D000440:Butanols; D004251:DNA Transposable Elements; D000096:Acetone; D000431:Ethanol; D014994:Xylose
keywords: Clostridium; random mutagenesis; transposon; xylose-inducible promoters