Synchronization of Caulobacter Crescentus for Investigation of the Bacterial Cell Cycle

Jared M. Schrader, Lucy Shapiro
2015 Journal of Visualized Experiments  
The cell cycle is important for growth, genome replication, and development in all cells. In bacteria, studies of the cell cycle have focused largely on unsynchronized cells making it difficult to order the temporal events required for cell cycle progression, genome replication, and division. Caulobacter crescentus provides an excellent model system for the bacterial cell cycle whereby cells can be rapidly synchronized in a G0 state by density centrifugation. Cell cycle synchronization
more » ... ts have been used to establish the molecular events governing chromosome replication and segregation, to map a genetic regulatory network controlling cell cycle progression, and to identify the establishment of polar signaling complexes required for asymmetric cell division. Here we provide a detailed protocol for the rapid synchronization of Caulobacter NA1000 cells. Synchronization can be performed in a large-scale format for gene expression profiling and western blot assays, as well as a small-scale format for microscopy or FACS assays. The rapid synchronizability and high cell yields of Caulobacter make this organism a powerful model system for studies of the bacterial cell cycle. Video Link The video component of this article can be found at
doi:10.3791/52633 pmid:25938623 pmcid:PMC4541484 fatcat:nzqzmvgm6jbubckqx4hl4btu7i