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PubMed · 15843930

Multiple large segment deletion method for Corynebacterium glutamicum.

Nobuaki Suzuki·Hiroshi Nonaka·Yota Tsuge·Satoshi Okayama·Masayuki Inui·Hideaki Yukawa
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Abstract

A precise and scarless genome excision method, employing the Cre/loxP system in concert with double-strand break (DSB)-stimulated intramolecular recombination was developed. The DSBs were mediated by the restriction endonuclease, I-SceI. It permitted multiple deletions of independent 14-, 43-, and 10-kb-long genomic regions on the Corynebacterium glutamicum genome. Accuracy of deletion was confirmed by the loss of marker genes, PCR, and sequencing of new genome joints. Eleven, 58, and 4 genes were predicted on the 14-, 43-, and 10-kb deleted regions, respectively. Although the resultant mutant lost a total of 67 kb encoding 73 genes, it still exhibited normal growth under standard laboratory conditions. Such a large segment deletion method in which multiple, successive deletions are possible is useful for genome engineering.

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Chromosome DeletionCorynebacterium glutamicumGenetic EngineeringGenome, BacterialSequence DeletionSubstrate SpecificityJournal ArticleResearch Support, Non-U.S. Gov't

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Nobuaki Suzuki, Hiroshi Nonaka, Yota Tsuge, Satoshi Okayama, Masayuki Inui, Hideaki Yukawa. 2005-11-12. Multiple large segment deletion method for Corynebacterium glutamicum.. https://doi.org/10.1007/s00253-005-1976-4

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