Improved response time
Estimate GFP-ssrA that can be degraded by ClpXP/ClpAP in molecules / min
Compare to reasonable detection limit in FACS
- Use pBAD UBP1 to modulate degradation rate
How to measure degradation rate below detection level? - can't be fluorescence!!!
- Western or pulse/chase IP
Do the same for PEST / proteasome in S.cerevisiae
Critical parameters for plasmid counting
Plasmid loss rate (when using ts plasmid)
- synthesis rate
- degradation rate
- maturation rate
How do these vary with temperature, growth media...