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Recombineering using RecET in Corynebacterium glutamicum ATCC14067 via a self-excisable cassette

Authors :
Nannan Zhao
Yuanyuan Huang
Ying Lin
Lu Li
Shan Xie
Shuangyan Han
Suiping Zheng
Source :
Scientific Reports, Vol 7, Iss 1, Pp 1-8 (2017), Scientific Reports
Publication Year :
2017
Publisher :
Nature Portfolio, 2017.

Abstract

Gene manipulation is essential for metabolic engineering and synthetic biology, but the current general gene manipulation methods are not applicable to the non-model strain Corynebacterium glutamicum (C. glutamicum) ATCC14067, which is used for amino acid production. Here, we report an effective and sequential deletion method for C. glutamicum ATCC14067 using the exonuclease-recombinase pair RecE + RecT (RecET) for recombineering via a designed self-excisable linear double-strand DNA (dsDNA) cassette, which contains the Cre/loxP system, to accomplish markerless deletion. To the best of our knowledge, this is the first effective and simple strategy for recombination with markerless deletion in C. glutamicum ATCC14067. This strategy provides a simple markerless deletion strategy for C. glutamicum and builds a solid basis for producer construction.

Details

Language :
English
ISSN :
20452322
Volume :
7
Issue :
1
Database :
OpenAIRE
Journal :
Scientific Reports
Accession number :
edsair.doi.dedup.....93ac270403812d226d2ffb9015e317ae