Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 80, Issue 1, Pages 106-108Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2009.10.013
Keywords
Francisella; Allelic replacement; Molecular genetics
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Funding
- National Institutes of Health [A1074402, T32 A1060525]
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [T32AI060525, R21AI074402] Funding Source: NIH RePORTER
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We engineered an efficient system to make Francisella tularensis deletion mutations using an unstable, poorly maintained plasmid to enhance the likelihood of homologous recombination. For counterselection, we adapted a strategy using I-SceI, which causes a double-stranded break in the integrated suicide vector, forcing a second recombination to mediate allelic replacement. (C) 2009 Elsevier B.V. All rights reserved.
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