Abstract
Mycobacteriophage L5, a temperate phage of the mycobacteria, forms stable lysogens in Mycobacterium smegmatis via site-specific integration of the phage genome. Recombination occurs within specific phage and bacterial attachment sites and is catalyzed by the phage-encoded integrase protein in vivo. We describe here the overexpression and purification of L5 integrase and its ability to mediate integrative recombination in vitro. We find that L5 integrase-mediated recombination is greatly stimulated by extracts of M. smegmatis but not by Escherichia coli extracts, purified E. coli integration host factor, or purified HU, indicating the presence of a novel mycobacterial integration host factor.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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DNA Nucleotidyltransferases / biosynthesis
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DNA Nucleotidyltransferases / isolation & purification
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DNA Nucleotidyltransferases / metabolism*
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DNA Primers
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Integrases
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Kinetics
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Molecular Sequence Data
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Mycobacteriophages / enzymology*
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Mycobacteriophages / genetics
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Mycobacterium / enzymology*
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Mycobacterium / genetics
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Plasmids
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Recombination, Genetic
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Restriction Mapping
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Thermodynamics
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Virus Integration*
Substances
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DNA Primers
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Recombinant Proteins
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DNA Nucleotidyltransferases
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Integrases