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.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Boudehen YM,Wallat M,Rousseau P,Neyrolles O,Gutierrez Cdoi
10.2144/btn-2019-0119subject
Has Abstractpub_date
2020-02-01 00:00:00pages
106-110issue
2eissn
0736-6205issn
1940-9818journal_volume
68pub_type
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