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  • Reconstitution of an efficient thymidine salvage pathway in Saccharomyces cerevisiae
    Vernis, Laurence ; Piškur, Jure, 1960-2014 ; Diffley, John F. X.
    The budding yeast Saccharomyces cerevisiae is unable to incorporate exogenous nucleosides into DNA. We have made a number of improvements to existing strategies to reconstitute an efficient thymidine ... salvage pathway in yeast. Wehave constructed strains that express both a nucleoside kinase as well as anequilibrative nucleoside transporter. By also deleting the gene encoding thymidylate synthase (CDC21) we have constructed strains that are entirely dependent upon exogenous thymidine for viability and that can grow with normalkinetics at low thymidine concentrations. Using this novel approach, we show that depletion of a single deoxyribonucleoside causes reversible arrest of cells in S phase with concomitant phosphorylation and activation of the S phase checkpoint kinase, Rad53. We show that this strain also efficiently incorporates the thymidine analogue, BrdU, into DNA and can be used for pulse-chase labelling.
    Vir: Nucleic acids research. - ISSN 0305-1048 (Vol. 31, no. 19, 2003, str. e120 [1-7])
    Vrsta gradiva - članek, sestavni del
    Leto - 2003
    Jezik - angleški
    COBISS.SI-ID - 25431257

vir: Nucleic acids research. - ISSN 0305-1048 (Vol. 31, no. 19, 2003, str. e120 [1-7])
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