An improved Xer-cise technology for the generation of multiple unmarked mutants in Mycobacteria - Archive ouverte HAL Access content directly
Journal Articles Biotechniques Year : 2020

An improved Xer-cise technology for the generation of multiple unmarked mutants in Mycobacteria

(1) , (1) , (2, 3) , (1) , (1)
1
2
3

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.

Dates and versions

hal-03019205 , version 1 (23-11-2020)

Identifiers

Cite

Yves-Marie Boudehen, Maximilian Wallat, Philippe Rousseau, Olivier Neyrolles, Claude Gutierrez. An improved Xer-cise technology for the generation of multiple unmarked mutants in Mycobacteria. Biotechniques, 2020, 68 (2), pp.106-110. ⟨10.2144/btn-2019-0119⟩. ⟨hal-03019205⟩
9 View
0 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More