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Site-specific integration of mycobacteriophage L5: Integration-proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille Calmette-Guerin

Mong Hong Lee, and Pascopella, L and Jacobs, WR and Hatfull, GF (1991) Site-specific integration of mycobacteriophage L5: Integration-proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille Calmette-Guerin. Proceedings of the National Academy of Sciences of the United States of America, 88 (8). 3111 - 3115. ISSN 0027-8424

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Abstract

Mycobacteriophage L5, a temperate phage of mycobacteria, integrates site- specifically into the Mycobacterium smegmatis chromosome. We have identified the int gene and attP site of L5, characterized the chromosomal attachment site (attB), and constructed plasmid vectors that efficiently transform M. smegmatis through stable site-specific integration of the plasmid into the bacterial genome. These integration-proficient plasmids also efficiently transform slow-growing mycobacteria such as the pathogen Mycobacterium tuberculosis and the vaccine strain bacille Calmette-Guerin (BCG). The ability to easily generate stable recombinants in these slow-growing mycobacteria without the requirement for continual selection is of particular importance for the construction of recombinant BCG vaccines and for the isolation and characterization of mycobacterial pathogenic determinants in animal model systems. Integration vectors of this type should be of general use in a number of additional bacterial systems where temperate phages have been identified.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Mong Hong Lee,
Pascopella, L
Jacobs, WR
Hatfull, GFgfh@pitt.eduGFH
Date: 1 January 1991
Date Type: Publication
Journal or Publication Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 88
Number: 8
Page Range: 3111 - 3115
DOI or Unique Handle: 10.1073/pnas.88.8.3111
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0027-8424
MeSH Headings: Amino Acid Sequence; Bacteriophages--genetics; Base Sequence; DNA, Bacterial--genetics; DNA, Viral--genetics; Genes, Viral; Genetic Vectors; Molecular Sequence Data; Mycobacterium--genetics; Mycobacterium bovis--genetics; Mycobacterium tuberculosis--genetics; Plasmids; Recombination, Genetic; Restriction Mapping
Other ID: NLM PMC51395
PubMed Central ID: PMC51395
PubMed ID: 1901654
Date Deposited: 12 Mar 2013 19:03
Last Modified: 02 Aug 2020 13:58
URI: http://d-scholarship.pitt.edu/id/eprint/17632

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