Septin (CDC10-YFP) marks bud neck and disappears at cytokinesis; tubulin (TUB1-GFP) marks spindle; elongates at anaphase and disassembles at telophase.
Driving synchronized cell cycles with periodic pulses of the anaphase inducer Cdc20.
The essential gene CDC20 is placed under control of the galactose inducible GAL1 promoter. The microfluidics flow-cell device of Charvin et al.( PLoS ONE. 2008 Jan 23;3(1):e1468) was used to provide periodic pulses of galactose at a period sufficient for each blocked cell to complete its cell cycle and reaccumulate at a new cdc20-deficient block. Green: tubulin (TUB1-YFP) marking mitotic spindle; yellow: CDC10-YFP: marker of bud neck, appears at budding and disappears at cytokinesis. Bottom: bars indicate periods of induction with galactose.

Periodic Start driven in a flow cell
Cells deleted for all G1 cyclins CLN1, CLN2 and CLN3 but provided with a methionine-regulatable MET3-CLN2 gene were periodically pulsed with methionine-free medium in the microfluidics flow-cell device of Charvin et al.( PLoS ONE. 2008 Jan 23;3(1):e1468). Green: Whi5-GFP; G1/S transcriptional repressor. Yellow: CDC10-YFP: marker of bud neck, appears at budding and disappears at cytokinesis. Bottom: bars indicate periods of induction with -Met.

Wild-type budding yeast cell cycle
Septin (CDC10-YFP) marks bud neck and disappears at cytokinesis; tubulin (TUB1-GFP) marks spindle; elongates at anaphase and disassembles at telophase.

Copyright F. Cross, The Rockefeller University. Not for use without proper attribution