Journal of Microbiology and Biotechnology
The Korean Society for Microbiology and Biotechnology publishes the Journal of Microbiology and Biotechnology.

2016 ; Vol.26-4: 725~729

AuthorSang-Hyun Lee, Hyun Ju Kim, Yong-An Shin, Kyoung Heon Kim, Sang Jun Lee
Place of dutyDepartment of Biotechnology, Graduate School, Korea University, Seoul 02841, Republic of Korea,R&D Center, GS Caltex Corporation, Daejeon 34122, Republic of Korea
TitleSingle Crossover-Mediated Markerless Genome Engineering in Clostridium acetobutylicum
PublicationInfo J. Microbiol. Biotechnol.2016 ; Vol.26-4
AbstractA novel genome-engineering tool in Clostridium acetobutylicum was developed based on singlecrossover homologous recombination. A small-sized non-replicable plasmid, pHKO1, was designed for efficient integration into the C. acetobutylicum genome. The integrated pHKO1 plasmid backbone, which included an antibiotic resistance gene, can be excised in vivo by Flp recombinase, leaving a single flippase recognition target sequence in the middle of the targeted gene. Since the pSHL-FLP plasmid, the carrier of the Flp recombinase gene, employed the segregationally unstable pAMβ1 replicon, the plasmid was rapidly cured from the mutant C. acetobutylicum. Consequently, our method makes it easier to engineer C. acetobutylicum.
Full-Text
Key_wordClostridium acetobutylicum, single crossover, gene inactivation, markerless
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