Mycobacteriophage L5 integrase-mediated site-specific integration in vitro.
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.
Lee
M H
MH
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260.
Hatfull
G F
GF
D000595:Amino Acid Sequence; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D017931:DNA Primers; D019426:Integrases; D007700:Kinetics; D008969:Molecular Sequence Data; D009160:Mycobacteriophages / Q000201:enzymology* / Q000235:genetics; D009161:Mycobacterium / Q000201:enzymology* / Q000235:genetics; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013816:Thermodynamics; D016662:Virus Integration
D017931:DNA Primers; D011994:Recombinant Proteins; D004254:DNA Nucleotidyltransferases; D019426:Integrases