Recombineering in Mycobacterium tuberculosis View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2007-02

AUTHORS

Julia C van Kessel, Graham F Hatfull

ABSTRACT

Genetic dissection of M. tuberculosis is complicated by its slow growth and its high rate of illegitimate recombination relative to homologous DNA exchange. We report here the development of a facile allelic exchange system by identification and expression of mycobacteriophage-encoded recombination proteins, adapting a strategy developed previously for recombineering in Escherichia coli. Identifiable recombination proteins are rare in mycobacteriophages, and only 1 of 30 genomically characterized mycobacteriophages (Che9c) encodes homologs of both RecE and RecT. Expression and biochemical characterization show that Che9c gp60 and gp61 encode exonuclease and DNA-binding activities, respectively, and expression of these proteins substantially elevates recombination facilitating allelic exchange in both M. smegmatis and M. tuberculosis. Mycobacterial recombineering thus provides a simple approach for the construction of gene replacement mutants in both slow- and fast-growing mycobacteria. More... »

PAGES

147-152

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nmeth996

DOI

http://dx.doi.org/10.1038/nmeth996

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1021371127

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/17179933


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