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 language | English |
|---|---|
| Pages (from-to) | 725-729 |
| Number of pages | 5 |
| Journal | Journal of Microbiology and Biotechnology |
| Volume | 26 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 2016 Apr 28 |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Single crossover-mediated markerless genome engineering in clostridium acetobutylicum
AU - Lee, Sang Hyun
AU - Kim, Hyun Ju
AU - Shin, Yong An
AU - Kim, Kyoung Heon
AU - Lee, Sang Jun
PY - 2016/4/28
Y1 - 2016/4/28
N2 - 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.
AB - 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.
KW - Clostridium acetobutylicum
KW - Gene inactivation
KW - Markerless
KW - Single crossover
UR - http://www.scopus.com/inward/record.url?scp=84964211353&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84964211353&partnerID=8YFLogxK
U2 - 10.4014/jmb.1512.12012
DO - 10.4014/jmb.1512.12012
M3 - Article
C2 - 26767573
AN - SCOPUS:84964211353
VL - 26
SP - 725
EP - 729
JO - Journal of Microbiology and Biotechnology
JF - Journal of Microbiology and Biotechnology
SN - 1017-7825
IS - 4
ER -