1887

Abstract

Mycobacteriophages provide an abundance of systems for use in mycobacterial genetics, including manipulation of . Because of the dearth of antibiotic resistance cassettes and biosafety concerns in constructing recombinant virulent strains, we developed the use of mycobacteriophage-encoded repressor genes that can be selected in the presence of lytic versions of their cognate phages. The phage Adephagia repressor gene () was identified through its ability to confer immunity to Adephagia superinfection, together with the mapping of mutations in gene that confer a clear-phage phenotype. Plasmid transformants containing either Adephagia or the previously identified BPs repressor can be readily selected following electroporation using engineered lytic derivatives of Adephagia and BPs, respectively. Selection is as efficient as antibiotic selection, can be used with either single-copy integration vectors or with extrachromosomal vectors, and works similarly in both and .

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2015-08-01
2019-12-13
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