Superinfection immunity of mycobacteriophage L5: applications for genetic transformation of mycobacteria |
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| Authors: | Mary K Donnelly-Wu William R Jacobs Jr Graham F Hatfull |
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| Institution: | Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.;Department of Microbiology and Immunology, Albert Einstein College of Medicine, Yeshiva University, 1300 Morris Park Avenue, Bronx, New York, New York 10461, USA. |
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| Abstract: | Mycobacteriophage L5 is a temperate phage of the mycobacteria that forms stable lysogens in Mycobacterium smegmatis. We show here that the 183-amino-acid product of L5 gene 71 confers immunity to L5 superinfection, is required for maintenance of the lysogenic state and contains a helix-turn-helix DNA-binding motif—properties associated with repressors of temperate phages. We have utilized these observations to demonstrate the use of L5 gene 71 as a selectable marker for genetic transformation of the mycobacteria. Significantly, the use of L5 gene 71 as a selectable gene avoids the requirement for antibiotic-resistance genes providing an important tool for manipulation of the pathogens Mycobacterium tuberculosis and Mycobacterium avium, and for the construction of recombinant BCG vaccines. |
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