Team:MIT/Experiments/Repressors

Testing Repressors

Purpose

This experiment aimed to produce and compare activity of the repressors BM3R1, TAL14, and TAL21.

Set Up

We created a plasmid containing each repressor under the control of a doxycycline inducible promoter, TRE. This allowed for us to modulate the repressor expression by varying the concentrations of doxycycline added. We also created a repressible promoter controlling the expression of a fluorescent protein. This plasmid allowed for us to measure the repression occurring.

Results

In order for us to analyze the level of repression, we need to see sustained presence of the repressor. Upon increasing the concentration of doxycycline, we were unable to see that sustained presence of the repressors. From the results of testing BM3R1 below, it can be seen that although there is activation early on, the production of repressor falls at 500 nM. This could possible be due to errors in doxycycline dilutions at 500 nM and above.