Construction of a pyrF Gene Mutant of Clostridium sp. AWRP Using the Clostron Mutagenesis System

DC Field Value Language
dc.contributor.author 채철기 -
dc.contributor.author 이종민 -
dc.contributor.author 도미숙 -
dc.contributor.author 권수재 -
dc.contributor.author 김윤재 -
dc.contributor.author 임재규 -
dc.contributor.author 강성균 -
dc.contributor.author 이현숙 -
dc.date.accessioned 2020-07-15T10:53:42Z -
dc.date.available 2020-07-15T10:53:42Z -
dc.date.created 2020-02-11 -
dc.date.issued 2018-10-04 -
dc.identifier.uri https://sciwatch.kiost.ac.kr/handle/2020.kiost/23105 -
dc.description.abstract Acetogenic bacteria uses the Wood-Ljungdahl pathway to fix either carbon monoxide (CO) or CO2 plus H2. These microorganisms have been receiving great attention for conversion of synthesis gas into industrially important feedstocks, including acetate, ethanol, and 2,3-butanediol (2,3-BDO). However, the typical production yield and titer obtained by these microorganisms are still low for economical production of such chemicals. Thus, metabolic engineering approaches are needed to further improve production performance. In this study, we developed basic genetic tools for Clostridium sp. AWRP, a newly isolated acetogenic species from a wetland at Ansan. As an example, we successfully disrupted the pyrF gene, encoding oritidine 5’-phosphate decarboxylase, in the AWRP strain using clostron mutagenesis. After transfer of the retargeted clostron plasmid via conjugation, the inactivation of the pyrF gene was both confirmed by erythromycin resistance and colony PCR. As expected, the pyrF mutants showed the resistance to 5-fluoroorotic acid (FOA), which becomes toxic after converted to 5-fluorouracil by pyrF. These results suggest that the tools developed in this study can be used for metabolic engineering and physiological studies on Clostridium sp. AWRP.g acetate, ethanol, and 2,3-butanediol (2,3-BDO). However, the typical production yield and titer obtained by these microorganisms are still low for economical production of such chemicals. Thus, metabolic engineering approaches are needed to further improve production performance. In this study, we developed basic genetic tools for Clostridium sp. AWRP, a newly isolated acetogenic species from a wetland at Ansan. As an example, we successfully disrupted the pyrF gene, encoding oritidine 5’-phosphate decarboxylase, in the AWRP strain using clostron mutagenesis. After transfer of the retargeted clostron plasmid via conjugation, the inactivation of the pyrF gene was both confirmed by erythromycin resistance and colony PCR. As expected, the pyrF mutants showed the resistance to 5-fluoroorotic acid (FOA), which becomes toxic after converted to 5-fluorouracil by pyrF. These results suggest that the tools developed in this study can be used for metabolic engineering and physiological studies on Clostridium sp. AWRP. -
dc.description.uri 1 -
dc.language English -
dc.publisher 한국해양과학기술원 -
dc.relation.isPartOf 2018 international Workshop on Deep-Sea Microbiology -
dc.title Construction of a pyrF Gene Mutant of Clostridium sp. AWRP Using the Clostron Mutagenesis System -
dc.type Conference -
dc.citation.conferenceDate 2018-10 -
dc.citation.conferencePlace KO -
dc.citation.title 2018 international Workshop on Deep-Sea Microbiology -
dc.contributor.alternativeName 채철기 -
dc.contributor.alternativeName 이종민 -
dc.contributor.alternativeName 도미숙 -
dc.contributor.alternativeName 권수재 -
dc.contributor.alternativeName 김윤재 -
dc.contributor.alternativeName 임재규 -
dc.contributor.alternativeName 강성균 -
dc.contributor.alternativeName 이현숙 -
dc.identifier.bibliographicCitation 2018 international Workshop on Deep-Sea Microbiology -
dc.description.journalClass 1 -
Appears in Collections:
Jeju Research Institute > Jeju Bio Research Center > 2. Conference Papers
Marine Resources & Environment Research Division > Marine Biotechnology &Bioresource Research Department > 2. Conference Papers
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