Efficient labeling of fission yeast Schizosaccharomyces pombe with thymidine and BUdR -: art. no. e134

被引:42
作者
Hodson, JA [1 ]
Bailis, JM [1 ]
Forsburg, SL [1 ]
机构
[1] Salk Inst Biol Studies, Mol & Cell Biol Lab, La Jolla, CA 92037 USA
关键词
D O I
10.1093/nar/gng134
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 H-3-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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页数:8
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共 37 条
  • [1] CLONING AND MANIPULATION OF THE SCHIZOSACCHAROMYCES-POMBE HIS7+ GENE AS A NEW SELECTABLE MARKER FOR MOLECULAR-GENETIC STUDIES
    APOLINARIO, E
    NOCERO, M
    JIN, M
    HOFFMAN, CS
    [J]. CURRENT GENETICS, 1993, 24 (06) : 491 - 495
  • [2] THYMIDINE 5'-MONOPHOSPHATE-REQUIRING MUTANTS OF SACCHAROMYCES-CEREVISIAE ARE DEFICIENT IN THYMIDYLATE SYNTHETASE
    BISSON, L
    THORNER, J
    [J]. JOURNAL OF BACTERIOLOGY, 1977, 132 (01) : 44 - 50
  • [3] Conserved organization of centromeric chromatin in flies and humans
    Blower, MD
    Sullivan, BA
    Karpen, GH
    [J]. DEVELOPMENTAL CELL, 2002, 2 (03) : 319 - 330
  • [4] Genetic analysis of an ARS element from the fission yeast Schizosaccharomyces pombe
    Clyne, RK
    Kelly, TJ
    [J]. EMBO JOURNAL, 1995, 14 (24) : 6348 - 6357
  • [5] Large, complex modular structure of a fission yeast DNA replication origin
    Dubey, DD
    Kim, SM
    Todorov, IT
    Huberman, JA
    [J]. CURRENT BIOLOGY, 1996, 6 (04) : 467 - 473
  • [6] The best yeast?
    Forsburg, SL
    [J]. TRENDS IN GENETICS, 1999, 15 (09) : 340 - 344
  • [7] Making sense of eukaryotic DNA replication origins
    Gilbert, DM
    [J]. SCIENCE, 2001, 294 (5540) : 96 - 100
  • [8] Organization of DNA replication origins in the fission yeast genome
    Gómez, M
    Antequera, F
    [J]. EMBO JOURNAL, 1999, 18 (20) : 5683 - 5690
  • [9] Cloning of a human nucleoside transporter implicated in the cellular uptake of adenosine and chemotherapeutic drugs
    Griffiths, M
    Beaumont, N
    Yao, SYM
    Sundaram, M
    Boumah, CE
    Davies, A
    Kwong, FYP
    Coe, I
    Cass, CE
    Young, JD
    Baldwin, SA
    [J]. NATURE MEDICINE, 1997, 3 (01) : 89 - 93
  • [10] THYMIDINE KINASE - EVIDENCE FOR ITS ABSENCE FROM NEUROSPORA CRASSA AND SOME OTHER MICRO-ORGANISMS AND RELEVANCE OF THIS TO SPECIFIC LABELLING OF DEOXYRIBONUCLEIC ACID
    GRIVELL, AR
    JACKSON, JF
    [J]. JOURNAL OF GENERAL MICROBIOLOGY, 1968, 54 : 307 - +