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Nucleic Acids Research, 2003, Vol. 31, No. 21 e134
© 2003 Oxford University Press

Efficient labeling of fission yeast Schizosaccharomyces pombe with thymidine and BUdR

Jeffrey A. Hodson, Julie M. Bailis and Susan L. Forsburg*

Molecular and Cell Biology Laboratory, The Salk Institute, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA

*To whom correspondence should be addressed. Tel: +1 858 453 4100; Fax: +1 858 457 4765; Email: forsburg{at}salk.edu

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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