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Identification and characterization of mycobacteriophage L5 excisionase

Lewis, JA and Hatfull, GF (2000) Identification and characterization of mycobacteriophage L5 excisionase. Molecular Microbiology, 35 (2). 350 - 360. ISSN 0950-382X

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Abstract

The well-characterized mycobacteriophage L5 forms stable lysogens in Mycobacterium smegmatis. Establishment of lysogeny involves integration of the phage genome into the chromosome of its mycobacterial hosts through an integrase-mediated site-specific recombination event. As L5 lysogens spontaneously generate free phage particles, prophage excision must also occur, although an L5 excisionase gene had not been identified. We show here that L5 gene 36 encodes the phage excisionase and is a small, heat-stable 56-amino-acid protein that strongly stimulates excisive recombination both in vivo and in vitro. The ability to manipulate the highly directional phage integration and excision reactions will provide powerful tools for the introduction, curing and recovery of foreign genes in recombinant mycobacterial strains.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Lewis, JA
Hatfull, GFgfh@pitt.eduGFH
Date: 31 January 2000
Date Type: Publication
Journal or Publication Title: Molecular Microbiology
Volume: 35
Number: 2
Page Range: 350 - 360
DOI or Unique Handle: 10.1046/j.1365-2958.2000.01695.x
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0950-382X
MeSH Headings: Amino Acid Sequence; Attachment Sites, Microbiological--genetics; DNA Nucleotidyltransferases--chemistry; DNA Nucleotidyltransferases--genetics; DNA Nucleotidyltransferases--metabolism; DNA, Viral--analysis; DNA, Viral--genetics; Electrophoresis, Polyacrylamide Gel; Molecular Sequence Data; Mycobacteriophages--enzymology; Mycobacteriophages--genetics; Mycobacteriophages--physiology; Mycobacterium smegmatis--virology; Plasmids--genetics; Polymerase Chain Reaction; Recombination, Genetic; Sequence Alignment; Viral Proteins; Virus Integration
PubMed ID: 10652095
Date Deposited: 29 Nov 2012 21:02
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16352

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