Single crossover-mediated markerless genome engineering in clostridium acetobutylicum

Sang Hyun Lee, Hyun Ju Kim, Yong An Shin, Kyoung Heon Kim, Sang Jun Lee

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

A 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.

Original languageEnglish
Pages (from-to)725-729
Number of pages5
JournalJournal of Microbiology and Biotechnology
Volume26
Issue number4
DOIs
Publication statusPublished - 2016 Apr 28

Fingerprint

Clostridium acetobutylicum
Plasmids
Genome
Recombinases
Genes
Replicon
Homologous Recombination
Microbial Drug Resistance

Keywords

  • Clostridium acetobutylicum
  • Gene inactivation
  • Markerless
  • Single crossover

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Single crossover-mediated markerless genome engineering in clostridium acetobutylicum. / Lee, Sang Hyun; Kim, Hyun Ju; Shin, Yong An; Kim, Kyoung Heon; Lee, Sang Jun.

In: Journal of Microbiology and Biotechnology, Vol. 26, No. 4, 28.04.2016, p. 725-729.

Research output: Contribution to journalArticle

Lee, Sang Hyun ; Kim, Hyun Ju ; Shin, Yong An ; Kim, Kyoung Heon ; Lee, Sang Jun. / Single crossover-mediated markerless genome engineering in clostridium acetobutylicum. In: Journal of Microbiology and Biotechnology. 2016 ; Vol. 26, No. 4. pp. 725-729.
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