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- Title
Development of a markerless gene replacement system in Corynebacterium glutamicum using upp as a counter-selection marker.
- Authors
Ma, Wenwen; Wang, Xiaoyue; Mao, Yufeng; Wang, Zhiwen; Chen, Tao; Zhao, Xueming
- Abstract
Corynebacterium glutamicum is well-established for industrial and biotechnological applications. However, its genetic manipulation has generally lagged behind traditional genetic models. In this study, a counter-selectable marker gene upp was firstly confirmed to be more efficient than traditional sacB. Furthermore, a markerless gene replacement system was developed by combining upp with double-strand break repair caused by the exogenous endonuclease I-S ceI. Finally, genetic modification using a dsDNA PCR fragment was carried out with the expression of recombinase/exonuclease RecE/RecT. Our results show that the genetic modification system allows precise and markerless gene replacement without altering the chromosome, with a simplified screening procedure to generate its modification.
- Subjects
CORYNEBACTERIUM glutamicum; GENE replacement; PHOSPHORIBOSYLTRANSFERASES; URACIL; DNA restriction enzymes; GENETIC recombination
- Publication
Biotechnology Letters, 2015, Vol 37, Issue 3, p609
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-014-1718-8