Utilization of an unstable plasmid and the I-SceI endonuclease to generate routine markerless deletion mutants in Francisella tularensis

作者：Horzempa Joseph; Shanks Robert M Q; Brown Matthew J; Russo Brian C; O'Dee Dawn M; Nau Gerard J^*

来源：Journal of Microbiological Methods, 2010, 80(1): 106-108.

DOI：10.1016/j.mimet.2009.10.013

• Francisella
• Allelic replacement
• Molecular genetics

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.

• 出版日期2010-1