Synchronization of Cell Cycles
From 2006.igem.org
Contents |
Group II
Aim
Our aim is to develop a synthetic gene network to synchronize E. coli cell cycles. Cell cycle is a robust biological oscillator. We plan to couple these oscillator using LuxI-LuxR based quorum sensing signaling. The cell cycle phase will be modulated using DnaA sequestration.
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.
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.
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)
