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Mycobacteriophage Bxb1 integrates into the Mycobacterium smegmatis groEL1 gene

Kim, AI and Ghosh, P and Aaron, MA and Bibb, LA and Jain, S and Hatfull, GF (2003) Mycobacteriophage Bxb1 integrates into the Mycobacterium smegmatis groEL1 gene. Molecular Microbiology, 50 (2). 463 - 473. ISSN 0950-382X

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Abstract

Mycobacteriophage Bxb1 is a temperate phage of Mycobacterium smegmatis and forms stable lysogens in which the Bxb1 genome is integrated into the host chromosome. Bxb1 encodes an integrase of the large serine recombinase family that catalyses integration and excision of the Bxb1 genome. We show here that Bxb1 integrates into a chromosomal attB site located within the 3' end of the groEL1 gene such that integration results in alteration of the C-terminal 21 amino acid residues. An integration-proficient plasmid vector containing the Bxb1 integrase gene and flanking DNA sequences efficiently transforms M. smegmatis via integration at attB. Bxb1-integrated recombinants are stable and fully compatible with L5 integration vectors. Strand exchange occurs within an 8 bp common core sequence present in attB and within an attP site situated immediately upstream of the phage integrase gene. Establishment of a defined in vitro system for Bxb1 integration shows that recombination occurs efficiently without requirement for high-energy cofactors, divalent metals, DNA supercoiling or additional proteins.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Kim, AI
Ghosh, P
Aaron, MA
Bibb, LA
Jain, S
Hatfull, GFgfh@pitt.eduGFH
Date: 1 October 2003
Date Type: Publication
Journal or Publication Title: Molecular Microbiology
Volume: 50
Number: 2
Page Range: 463 - 473
DOI or Unique Handle: 10.1046/j.1365-2958.2003.03723.x
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0950-382X
MeSH Headings: Amino Acid Sequence; Chaperonin 60--genetics; DNA, Bacterial--genetics; DNA, Viral--genetics; Genes, Viral; Genetic Vectors; Lysogeny; Molecular Sequence Data; Mycobacteriophages--genetics; Mycobacterium--genetics; Plasmids; Repressor Proteins--genetics; Viral Proteins--genetics; Virus Integration
PubMed ID: 14617171
Date Deposited: 29 Nov 2012 21:04
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16323

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