1887

Abstract

The R1 genome encodes an X-family DNA repair polymerase homologous to eukaryotic DNA polymerase . The recombinant deinococcal polymerase X (PolX) purified from transgenic showed deoxynucleotidyltransferase activity. Unlike the Klenow fragment of , this enzyme showed short patch DNA synthesis activity on heteropolymeric DNA substrate. The recombinant enzyme showed 5′-deoxyribose phosphate (5′-dRP) lyase activity and base excision repair function with the help of externally supplied glycosylase and AP endonuclease functions. A disruption mutant of expressing 5′-dRP lyase and a truncated polymerase domain was comparatively less sensitive to -radiation than a deletion mutant. Both mutants showed higher sensitivity to hydrogen peroxide. Excision repair mutants of expressing this polymerase showed functional complementation of UV sensitivity. These results suggest the involvement of deinococcal polymerase X in DNA-damage tolerance of , possibly by contributing to DNA double-strand break repair and base excision repair.

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2009-09-01
2019-12-07
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vol. , part 9, pp. 3005 - 3014

[ PDF, 150 kb], including: Characterization of pPolXcat plasmid used for generating the disruptant mutant of Construction of pNOKpolX plasmid used for generating the deletion mutant of Translation of the : gene for checking the disruption of the translation reading frame by insertion Cloning of in pET28a+ and inducible expression of recombinant polymerase X in Bacterial strains used Primers used



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