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Recombineering mycobacteria and their phages

Nature Reviews Microbiology volume 6, pages 851–857 (2008)Cite this article

Abstract

Bacteriophages are central components in the development of molecular tools for microbial genetics. Mycobacteriophages have proven to be a rich resource for tuberculosis genetics, and the recent development of a mycobacterial recombineering system based on mycobacteriophage Che9c-encoded proteins offers new approaches to mycobacterial mutagenesis. Expression of the phage exonuclease and recombinase substantially enhances recombination frequencies in both fast- and slow-growing mycobacteria, thereby facilitating construction of both gene knockout and point mutants; it also provides a simple and efficient method for constructing mycobacteriophage mutants. Exploitation of host-specific phages thus provides a general strategy for recombineering and mutagenesis in genetically naive systems.

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Figure 1: Mycobacteriophage-encoded recombination proteins.
Figure 2: Strategies for mycobacterial recombineering.
Figure 3: Mutagenesis of mycobacterial genomes.
Figure 4: Mycobacteriophage recombineering.

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Acknowledgements

This work was supported by US National Institutes of Health grant AI067649. We thank W. Jacobs Jr and D. Court for helpful discussions, and M. Bochman for help with electron microscopy.

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Authors and Affiliations

  1. Laura J. Marinelli and Graham F. Hatfull are at the Pittsburgh Bacteriophage Institute and Department of Biological Sciences, Julia C. van Kessel, 376 Crawford Hall, 4249 Fifth Avenue, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.,

    Julia C. van Kessel, Laura J. Marinelli & Graham F. Hatfull

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  1. Julia C. van Kessel
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Correspondence to Graham F. Hatfull.

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DATABASES

Entrez Genome

Che9c

Halo

Giles

lambda

P22

TM4

Entrez Genome Project

Escherichia coli

Mycobacterium avium

Mycobacterium bovis BCG

Mycobacterium tuberculosis

Mycobacterium smegmatis

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van Kessel, J., Marinelli, L. & Hatfull, G. Recombineering mycobacteria and their phages. Nat Rev Microbiol 6, 851–857 (2008). https://doi.org/10.1038/nrmicro2014

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