Sulfolobus Mutants, Generated via PCR Products, Which Lack Putative Enzymes of UV Photoproduct Repair. (7th July 2011)

• Record Type: Journal Article
• Authors: Sakofsky, Cynthia J.; Runck, Laura A.; Grogan, Dennis W.
• Other Names: Ishino Y. Academic Editor.
• Is Part Of: Archaea. Volume 2011(2011)
• Publication Date: 2011-07-07
• Subjects: Archaebacteria -- Periodicals; 579.321
• DOI: 10.1155/2011/864015
• Languages: English
• ISSNs: 1472-3646

Abstract

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.