Synchronization of Cell Cycles

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Group II

Aim

We are interested in synthetically synchronizing cell cycles.

Approach

Constructs

In our genetic construct, we have used Pcya (adenylate cyclase promoter-that is known to oscillate in cell cycle dependant manner) as a core oscillator.

Testing oscillations

LuxI tagged with cfp(+LVA) is expressed from this promoter.LuxI oscillations will result in AI(autoinducer) oscillations and this would result in oscillations of AI dependant promoter. YFP is expressed from such a promoter. CFP expression is readout for the core oscillator-Pcya and YFP is readout of the expression of the quorum sensing promoter-Plux.

Testing the core oscillator Pcya

Y axis is the mean cfp fluorescence of a cell. X-axis is a correlate of cell cycle phase.Here, we have used cell length as a crude correlate for cell cycle phase.The same data is platted twice for visual representation of oscillation. RFP expression is control.

Testing oscillations at Plux

Y axis is the mean yfp fluorescence of a cell. X-axis is major axis length of a cell which is a correlate of cell cycle phase.

--Sugat 06:49, 8 July 2006 (EDT)