Adam Cohen’s PROPS sensor is a fluorescent indicator of the proton-motive force. Here’s an adaption of the protocol to induce expression of the PROPS protein (which is on the pBAD promoter).
Materials
- 20 mM all-trans retinal stock solution in ethanol
- 10% w/v arabinose stock
- 1X M9 salt with 0.4% glucose, pH 7.0
- LB
Procedure
- Grow strain in 50 mL of LB at 33C until you reach an OD = 0.3-0.4
- Add arabinose (100 uL of stock, i.e. to 0.02%) and all-trans retinal (12.5 uL of stock, i.e. to 5 uM)
- Grow in dark for 3.5 hours at 33C
- Harvest cells and wash in 30 mL of the M9 medium
- Resuspend cells in 5 mL M9 medium and store at 4C for later use (can store up to 2 weeks)