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An improved Xer-cise technology for the generation of multiple unmarked mutants in Mycobacteria
Abstract : Xer-cise is a technique using antibiotic resistance cassettes flanked by dif sites allowing spontaneous and accurate excision from bacterial chromosomes with a high frequency through the action of the cellular recombinase XerCD. Here, we report a significant improvement of Xer-cise in Mycobacteria. Zeocin resistance cassettes flanked by variants of the natural Mycobacterium tuberculosis dif site were constructed and shown to be effective tools to construct multiple unmarked mutations in M. tuberculosis and in the model species Mycobacterium smegmatis. The dif site variants harbor mutations in the central region and can therefore not recombine with the wild-type or other variants, resulting in mutants of increased genetic stability. The herein described method should be generalizable to virtually any transformable bacterial species.
https://hal-cnrs.archives-ouvertes.fr/hal-03019205
Contributor : Colette Orange Connect in order to contact the contributor
Submitted on : Monday, November 23, 2020 - 11:48:57 AM
Last modification on : Wednesday, November 3, 2021 - 6:36:59 AM
Contributor : Colette Orange Connect in order to contact the contributor
Submitted on : Monday, November 23, 2020 - 11:48:57 AM
Last modification on : Wednesday, November 3, 2021 - 6:36:59 AM
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