title: Sulfolobus mutants, generated via PCR products, which lack putative enzymes of UV photoproduct repair.
abstract: In order to determine the biological relevance of two S. acidocaldarius proteins to the repair of UV photoproducts, the corresponding genes (Saci_1227 and Saci_1096) were disrupted, and the phenotypes of the resulting mutants were examined by various genetic assays. The disruption used integration by homologous recombination of a functional but heterologous pyrE gene, promoted by short sequences attached to both ends via PCR. The phenotypic analyses of the disruptants confirmed that ORF Saci_1227 encodes a DNA photolyase which functions in vivo, but they could not implicate ORF Saci_1096 in repair of UV- or other externally induced DNA damage despite its similarity to genes encoding UV damage endonucleases. The success of the gene-disruption strategy, which used 5' extensions of PCR primers to target cassette integration, suggests potential advantages for routine construction of Sulfolobus strains.
authors.affiliation: Department of Biological Sciences, University of Cincinnati, 614 Rieveschl Hall, Clifton Court, Cincinnati, OH 45221-0006, USA.
authors.forename: Cynthia J
authors.initials: CJ
authors.lastname: Sakofsky
authors.forename: Laura A
authors.initials: LA
authors.lastname: Runck
authors.forename: Dennis W
authors.initials: DW
authors.lastname: Grogan
mesh_terms: D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000235:genetics*; D019641:DNA, Archaeal / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D005818:Genetic Engineering; D053858:Metabolic Networks and Pathways; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D016945:Sulfolobus acidocaldarius / Q000235:genetics* / Q000528:radiation effects; D014466:Ultraviolet Rays
chemical_list: D019641:DNA, Archaeal; D045643:DNA Repair Enzymes