title: Purification of the bacteriophage lambda xis gene product required for lambda excisive recombination.
abstract: Excision of the lambda prophage from the chromosome of its Escherichia coli host requires the products of the two viral genes int and xis. This paper reports a purification of the lambda xis gene product using a complementation assay in which functional Xis must be added to purified Int and an E. coli-derived host factor extract. Excisive recombination between a left (attL) and right (attR) prophage attachment site cloned on the same plasmid DNA substrate occurred efficiently under these conditions. Purified Int and Xis together could not carry out excision in vitro unless an extract derived from the E. coli host was added; purified integration host factor satisfied this requirement. Xis appears to have a molecular weight of 8800 as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. It possesses no detectable endonuclease or topoisomerase activities, does not appear to bind DNA to filters, and does not increase the ability of Int to bind DNA. The addition of Xis not only stimulated excisive recombination in vitro but also inhibited integrative recombination. Xis protected Int protein from heat inactivation, suggesting a possible interaction between the two proteins. In light of these observations, possible roles for Xis in recombination are discussed.
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics*; D002845:Chromatography; D004279:DNA, Viral / Q000378:metabolism; D004926:Escherichia coli / Q000032:analysis*; D005814:Genes, Viral; D007700:Kinetics; D010957:Plasmids; D011995:Recombination, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins