Hlavová Monika, Vítová Milada, Bišová Kateřina
Laboratory of Cell Cycles of Algae, Centre Algatech, Institute of Microbiology CAS, Opatovický mlýn, Třeboň, 379 81, Czech Republic.
Methods Mol Biol. 2016;1370:3-16. doi: 10.1007/978-1-4939-3142-2_1.
A synchronous population of cells is one of the prerequisites for studying cell cycle processes such as DNA replication, nuclear and cellular division. Green algae dividing by multiple fission represent a unique single cell system enabling the preparation of highly synchronous cultures by application of a light-dark regime similar to what they experience in nature. This chapter provides detailed protocols for synchronization of different algal species by alternating light-dark cycles; all critical points are discussed extensively. Moreover, detailed information on basic analysis of cell cycle progression in such cultures is presented, including analyses of nuclear, cellular, and chloroplast divisions. Modifications of basic protocols that enable changes in cell cycle progression are also suggested so that nuclear or chloroplast divisions can be followed separately.
细胞同步群体是研究诸如DNA复制、细胞核和细胞分裂等细胞周期过程的前提条件之一。通过多次分裂进行繁殖的绿藻代表了一种独特的单细胞系统,通过应用类似于它们在自然环境中所经历的明暗周期,能够制备出高度同步的培养物。本章提供了通过交替明暗循环使不同藻类物种同步化的详细方案;对所有关键点进行了广泛讨论。此外,还介绍了此类培养物中细胞周期进程基本分析的详细信息,包括细胞核、细胞和叶绿体分裂的分析。还提出了能够改变细胞周期进程的基本方案的修改建议,以便能够分别追踪细胞核或叶绿体的分裂。