Modeling and measurements for flipper operation
Detail Mechanism
- Inducer IPTG diffuse into a cell
- Mechanism: IPTG diffuses across E.coli cell membrane and cytosol and binds to lacI
- Rate
- Diffusion rate of IPTG across cell membrane
- Diffusion rate of IPTG in cytosol
- IPTG-lacI binding rate
- Number
- number of lacI in a single cell
- lacI-IPTG leaves the promoter
- Mechanism: lacI-IPTG has much lower affinity toward Lac operator than lacI and likely to dissociate
- Rate
- Dissociation rate of lacI-IPTG from the lac operon
- Number
- Transcription of Flipper
- Mechanism: RNAP bind to the promoter, transcribing mRNA of the flipper out
- Rate
- Estimated transcription rate
- promoter occupancy of our promoter
- Number
- Size of the gene
- number of RNAP molecules
- Translation of Flipper
- Flipper localizes at Flippee
- DNA inversion
- Transcription of reporter
- Translation of reporter
- Maturation of reporter
- Detection
Measurement
(Currently available in our lab)
- Bulk GFP signal readout
- Bulk b-gal activity readout
- Bulk Restriction Digestion
- Bulk PCR
- Single-cell GFP readout
(Not available in our lab yet)
- IPTG diffusion
- Bulk Flipper transcription
- Single-cell fipper transcription
- Bulk Flipper translation
- Single-cell flipper translation
- Flipper localizes on the flippee
- in vivo single cell DNA inversion
- Bulk Flippee transcription
- Single-cell Flippee translation
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