Efficient labeling of fission yeast Schizosaccharomyces pombe with thymidine and BUdR | Zendy

Jeffrey A. Hodson | Zendy

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Efficient labeling of fission yeast Schizosaccharomyces pombe with thymidine and BUdR

Author(s) -

Jeffrey A. Hodson

Publication year - 2003

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gng134

Subject(s) - schizosaccharomyces pombe , biology , thymidine , thymidine kinase , schizosaccharomyces , dna , uridine , dna replication , microbiology and biotechnology , yeast , gene , genetics , herpes simplex virus , saccharomyces cerevisiae , rna , virus

In this paper we report the construction of a Schizosaccharomyces pombe strain that facilitates analysis of replicating DNA. The strain co-expresses the Herpes simplex virus thymidine kinase gene (hsv-tk) and a human equilibrative nucleoside transporter (hENT1). The double integrant efficiently incorporates 3H-thymidine into nuclear DNA as monitored by scintillation counting. These strains also incorporate the thymidine analog Bromodeoxy uridine (BUdR) into newly replicated DNA, which can be detected by immunofluorescence and flow cytometry. This strain provides a valuable tool for direct study of DNA replication in S.pombe

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