Multiple large segment deletion method for Corynebacterium glutamicum
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.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2005 |
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| Erschienen: |
2005 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:69 |
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| Enthalten in: |
Applied microbiology and biotechnology - 69(2005), 2 vom: 15. Nov., Seite 151-61 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Suzuki, Nobuaki [VerfasserIn] |
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| Themen: |
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| Anmerkungen: |
Date Completed 08.08.2006 Date Revised 15.11.2006 published: Print-Electronic Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM154928518 |
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| 100 | 1 | |a Suzuki, Nobuaki |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Multiple large segment deletion method for Corynebacterium glutamicum |
| 264 | 1 | |c 2005 | |
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| 500 | |a Date Completed 08.08.2006 | ||
| 500 | |a Date Revised 15.11.2006 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 700 | 1 | |a Nonaka, Hiroshi |e verfasserin |4 aut | |
| 700 | 1 | |a Tsuge, Yota |e verfasserin |4 aut | |
| 700 | 1 | |a Okayama, Satoshi |e verfasserin |4 aut | |
| 700 | 1 | |a Inui, Masayuki |e verfasserin |4 aut | |
| 700 | 1 | |a Yukawa, Hideaki |e verfasserin |4 aut | |
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| 773 | 1 | 8 | |g volume:69 |g year:2005 |g number:2 |g day:15 |g month:11 |g pages:151-61 |
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