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

Hong Lee Mong Hong Lee, L. Pascopella, W. R. Jacobs, G. F. Hatfull

Research output: Contribution to journalArticle

287 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)3111-3115
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number8
DOIs
StatePublished - Jan 1 1991

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Keywords

  • Mycobacterium leprae
  • pathogenesis
  • site-specific recombination
  • vaccines

ASJC Scopus subject areas

  • General

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