Endy:Screening plasmid/v2.0/Testing

There are a few tests we can get started on before the complete SP2.0 is constructed.

=Induction system test=

AHL-induction test

 * I13546 (F2620 + GFP/RFP) on psb3K3
 * I13547 (F2621 + GFP/RFP) on psb3K3

Date?

 * 
 * Looks like the GFP is responding well over a range of AHL conentrations. So the induction system itself looks pretty good.  In fact the GFP expression was higher than we had seen with pBAD on a high copy (psb1A2) plasmid, and F262X were on 3k3.
 * However, the RFP is displaying low fluorecence intensity. Planning experiments to figure out why - see below.
 * Also, the data collection was set to linear scale, so will be redoing this experiment on 9/21/2006

9/21/2006

 * I13546 (F2620 + GFP/RFP) on psb3K3
 * Induced 1nM, 10nM, 50nM, 100nM, 500nM, 1uM
 * I13547 (F2621 + GFP/RFP) on psb3K3
 * Induced 1nM, 5nM, 10nM, 50nM, 100nM, 1uM
 * The variation in the induction levels was based on the results of the previous experiment, which demonstrated over which induction range the device was switching in response to AHL.

Date?

 * Will be comparing the expression of several costructs:
 * I13530 - pbad controlled RFP
 * I13521 - Ptet controlled RFP
 * I13549 - F2620 controlled RFP
 * I13550 - F2621 controlled RFP
 * I13551 - F2620 controlled RnaseE+hairpin+RFP
 * I13552 - F2621 controlled RnaseE+hairpin+RFP
 * All will be induced at the highest induction level (except tet which is constitive in this case because no tetR present).
 * The RFP lacking the RnaseE site and the hairpin produced a weaker signal in response to equivalent induction. This is counter to some of our previous results using the pBAD induction system.  Should consider revisiting those.
 * The RFP lacking the RnaseE site and the hairpin produced a weaker signal in response to equivalent induction. This is counter to some of our previous results using the pBAD induction system.  Should consider revisiting those.

=SP1.0 debugging= Understanding the issues with GFP/RFP relationship in SP1.0 will help us to better debug SP2.0 down the line.

Arabinose test
Will use a construct that has a constituitive promoter (R0074) driving expression of GFP/RFP. Will then grow in the presence of varying levels of arabinose and look at the effect on GFP/RFP levels.
 * I13553