ArticleMicrobiology spectrum2025
Induction-free recombineering for simple targeted gene-deletions in various mycobacteria.
Article in Microbiology spectrum, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Gene deletion is a valuable tool for phenotypic characterization in bacteriology, but in mycobacteria, generating deletion mutants remains a cumbersome and time-consuming process. Here, we present a modification to the widely used recombineering method in mycobacteria. By expressing the recombineering-promoting proteins from a single-copy episomal plasmid, we could use a constitutive promotor. This approach eliminates the need for induction prior to electroporation of the targeting substrate (linear DNA fragment comprised of homologous regions to the gene of interest, flanking a selection marker), and shortens the subsequent plasmid curing process. We successfully demonstrated this method in
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