Nielsen Olaf
Department of Biology, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
Cold Spring Harb Protoc. 2016 Sep 1;2016(9):2016/9/pdb.prot091272. doi: 10.1101/pdb.prot091272.
A well-characterized S phase, a unicellular lifestyle, and a plethora of mutations in key components of DNA metabolism make fission yeast a particularly attractive system in which to study DNA replication. However, synchronization of passage through a normal S phase has proved challenging. This protocol describes how combining nitrogen starvation with M-factor mating pheromone treatment presents a highly effective method for synchronizing passage through an ostensibly normal S phase.
特征明确的S期、单细胞生活方式以及DNA代谢关键成分中的大量突变,使得裂殖酵母成为研究DNA复制的极具吸引力的系统。然而,事实证明,使细胞同步通过正常S期具有挑战性。本方案描述了如何将氮饥饿与M因子交配信息素处理相结合,从而提供一种使细胞同步通过表面上正常S期的高效方法。