SulfolobusMutants, Generated via PCR Products, Which LackPutative Enzymes of UV Photoproduct Repair

[摘要] In order to determine the biological relevance of twoS. acidocaldariusproteins 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 heterologouspyrEgene, promoted by short sequences attached to both endsviaPCR. The phenotypic analyses of the disruptants confirmed that ORF Saci_1227 encodes a DNA photolyase which functionsin 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 ofSulfolobusstrains.

[学科分类] 微生物学和免疫学