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KMID : 0545120150250101702
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Journal of Microbiology and Biotechnology
2015 Volume.25 No. 10 p.1702 ~ p.1708
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A New Shuttle Plasmid That Stably Replicates in Clostridium acetobutylicum
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Lee Sang-Hyun
Kwon Min-A
Choi Sun-Hwa
Kim Soo-Ah
Kim Jung-Yeon
Shin Yong-An
Kim Kyoung-Heon
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Abstract
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We have developed a new shuttle plasmid, designated as pLK1-MCS that can replicate in both Clostridium acetobutylicum and Escherichia coli, by combining the pUB110 and pUC19 plasmids. Plasmid pLK1-MCS replicated more stably than previously reported plasmids containing either the pIM13 or the pAMβ1 replicon in the absence of antibiotic selective pressure. The transfer frequency of pLK1-MCS into C. acetobutylicum was similar to the transfer frequency of other shuttle plasmids. We complemented C. acetobutylicum ML1 (that does not produce solvents such as acetone, butanol, and ethanol owing to loss of the megaplasmid pSOL1 harboring the adhE1-ctfAB-adc operon) by introducing pLK1-MCS carrying the adhE1-ctfAB-adc operon into C. acetobutylicum ML1. The transformed cells were able to resume anaerobic solvent production, indicating that the new shuttle plasmid has the potential for practical use in microbial biotechnology.
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KEYWORD
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Clostridium acetobutylicum, shuttle plasmid, segregational stability
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