Tormos-Pérez Marta, Pérez-Hidalgo Livia, Moreno Sergio
Instituto de Biología Funcional y Genómica, Consejo Superior de Investigaciones Científicas/Universidad de Salamanca, 37007, Salamanca, Spain.
Instituto de Investigación Biomédica de Salamanca (IBSAL), 37007, Salamanca, Spain.
Methods Mol Biol. 2016;1369:293-308. doi: 10.1007/978-1-4939-3145-3_20.
Fission yeast cells can be synchronized by cell cycle arrest and release or by size selection. Cell cycle arrest synchronization is based on the block and release of temperature-sensitive cell cycle mutants or treatment with drugs. The most widely used approaches are cdc10-129 for G1; hydroxyurea (HU) for early S-phase; cdc25-22 for G2, and nda3-KM311 for mitosis. Cells can also be synchronized by size selection using centrifugal elutriation or a lactose gradient. Here we describe the methods most commonly used to synchronize fission yeast cells.
裂殖酵母细胞可以通过细胞周期阻滞与释放或通过大小选择来实现同步化。细胞周期阻滞同步化基于对温度敏感的细胞周期突变体的阻断与释放或药物处理。最广泛使用的方法是:用于G1期的cdc10-129;用于早S期的羟基脲(HU);用于G2期的cdc25-22,以及用于有丝分裂的nda3-KM311。细胞也可以使用离心淘析或乳糖梯度通过大小选择来实现同步化。在此,我们描述最常用于使裂殖酵母细胞同步化的方法。