title: A New Shuttle Plasmid That Stably Replicates in Clostridium acetobutylicum.
abstract: 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.
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul 136-713, Republic of Korea.|R&D Center, GS Caltex Corporation, Daejeon 305-380, Republic of Korea.
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Lee
authors.affiliation: R&D Center, GS Caltex Corporation, Daejeon 305-380, Republic of Korea.
authors.forename: Min-A
authors.initials: MA
authors.lastname: Kwon
authors.affiliation: R&D Center, GS Caltex Corporation, Daejeon 305-380, Republic of Korea.
authors.forename: Sunwha
authors.initials: S
authors.lastname: Choi
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Sooah
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Jungyeon
authors.initials: J
authors.lastname: Kim
authors.affiliation: R&D Center, GS Caltex Corporation, Daejeon 305-380, Republic of Korea.
authors.forename: Yong-An
authors.initials: YA
authors.lastname: Shin
authors.affiliation: Department of Biotechnology, Graduate School, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Kyoung Heon
authors.initials: KH
authors.lastname: Kim
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism; D046969:Clostridium acetobutylicum / Q000235:genetics*; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D042822:Genomic Instability; D060847:Metabolic Engineering / Q000379:methods; D053858:Metabolic Networks and Pathways / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids; D012641:Selection, Genetic; D012997:Solvents / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D012997:Solvents
keywords: Clostridium acetobutylicum; segregational stability; shuttle plasmid