Week 14

9/22/13

Today we set up another run identical to the one from yesterday in order to have duplicated data of pTet+TBS 2+ GFP+pBAD+RS1+TAL8. We will then average this data out and use it to understand natural deviations that may occur in our genetic circuit.

After measuring fluorescence of our cotransformation involving pTet+TBS 2+GFP and pBAD+RS1+TAL 8, we decided to do begin another run with varying concentrations of our inducers for pTet+TBS 2+GFP and pBAD+RS2+TAL8.

Instead of testing a gradient of different inducer concentrations, we decided to run the rest of our assembled constructs by adding three different amounts of arabinose and two different concentrations of theophylline. aTc was always added at 100 ng/mL for any construct involving Ptet. We did this to conserve space for triplicate runs to see the most extreme cases of repression. In this final run, we tested all the different combinations of TALs and riboswitches as well as Ptet+TBS 1+GFP and Ptet+TBS 2+GFP.

After looking at the data, it appears that the most extreme cases of induction are behaving as expected, but some intermediate concentrations of arabinose are showing only slight repression. Both of the riboswitches are also showing slightly different responses to induction, which is due to leakiness. Interestingly, Ptet+TBS 1+GFP and Ptet+TBS 2+GFP both exhibited different levels of GFP expression at full aTc induction. We are unsure as to what is the source of this discrepancy.

School started today, but we’re all hard at work on the Wiki. We’re refining the last of our data that we will use for our presentation, Wiki, and poster. In addition, we’ve been working on the modeling of our genetic circuits.

With the Wiki freeze today, we’re fine tuning our website. We’re also filling out all the parts characterization data for the registry. We still need to fill out the judging forms and hopefully all of this can be done by the end of the day. Almost finished!