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Characterization of the Mycobacteriophage L5 attachment site, attP

Peña, CEA and Lee, MH and Pedulla, ML and Hatfull, GF (1997) Characterization of the Mycobacteriophage L5 attachment site, attP. Journal of Molecular Biology, 266 (1). 76 - 92. ISSN 0022-2836

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Lysogenization of mycobacteriophage L5 involves integration of the phage genome into the Mycobacterium smegmatis chromosome. Integration occurs by a site-specific recombination event between a phage attachment site, attP, and a bacterial attachment site, attB, which is catalyzed by the phage-encoded integrase protein. DNase I footprinting reveals that L5 integrase binds to two types of sites within attP which span an unexpectedly large region of 413 bp: seven arm-type sites (P1 to P7) each of which correspond to a consensus sequence 5'-TGCaaCtcYy, and core-type sites at the points of strand exchange. Mutational analyses indicate that not all of the arm-type sites are required for integration, and that the P3 site and the rightmost pair of sites (P6 and P7) are dispensable for Integration. We show that a 252 bp segment of attP DNA is sufficient for efficient integrative recombination and that int can be provided in trans for simple and efficient transformation of the mycobacteria.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Peña, CEA
Lee, MH
Pedulla, ML
Hatfull, GFgfh@pitt.eduGFH
Date: 14 February 1997
Date Type: Publication
Journal or Publication Title: Journal of Molecular Biology
Volume: 266
Number: 1
Page Range: 76 - 92
DOI or Unique Handle: 10.1006/jmbi.1996.0774
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0022-2836
MeSH Headings: Bacteriophages--genetics; Bacteriophages--physiology; Base Sequence; Binding Sites; Chromosomes, Bacterial; Cloning, Molecular; Consensus Sequence; DNA Footprinting; DNA Mutational Analysis; Deoxyribonuclease I; Escherichia coli; Genome, Viral; Integrases--metabolism; Molecular Sequence Data; Mycobacterium--genetics; Mycobacterium--virology; Recombination, Genetic; Sequence Deletion; Virus Integration
PubMed ID: 9054972
Date Deposited: 12 Mar 2013 19:09
Last Modified: 02 Feb 2019 15:57


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