Cell-Cycle Analyses Using Thymidine Analogues in Fission Yeast

被引：14

作者：

Anda, Silje ^{[1
]}

Boye, Erik ^{[1
]}

Grallert, Beata ^{[1
]}

机构：

[1] Oslo Univ Hosp, Inst Canc Res, Dept Cell Biol, Oslo, Montebello, Norway

来源：

PLOS ONE | 2014年 / 9卷 / 02期

关键词：

SCHIZOSACCHAROMYCES-POMBE; DNA-REPLICATION; IN-VIVO; CHECKPOINT; REPAIR; MUTAGENESIS; DAMAGE;

D O I：

10.1371/journal.pone.0088629

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Thymidine analogues are powerful tools when studying DNA synthesis including DNA replication, repair and recombination. However, these analogues have been reported to have severe effects on cell-cycle progression and growth, the very processes being investigated in most of these studies. Here, we have analyzed the effects of 5-ethynyl-2'-deoxyuridine (EdU) and 5-Chloro-2'-deoxyuridine (CldU) using fission yeast cells and optimized the labelling procedure. We find that both analogues affect the cell cycle, but that the effects can be mitigated by using the appropriate analogue, short pulses of labelling and low concentrations. In addition, we report sequential labelling of two consecutive S phases using EdU and 5-bromo- 2'-deoxyuridine (BrdU). Furthermore, we show that detection of replicative DNA synthesis is much more sensitive than DNA-measurements by flow cytometry.

引用

页数：9

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