Research Status: Cloned, Sequenced and Ready for the Real Stuff
So because Wednesday was July 4th, this week has been slower than usual but nonetheless much was accomplished. First, on Friday Jack and I presented an article about TGF-B signalling in prostate cancer stromal cells for our lab’s biweekly journal club meeting. It was interesting to learn about what approaches other labs are doing to study TGF-B. We did another Western blot to detect if our ”protein X” fragments were successfully expressed. However, since we still didn’t detect any signal for the fragments of interest, we immunoprecipitated the fragments with an antibody before doing the Western to reduce any nonspontaneous proteins binding to the membrane. As far as the mutagenesis experiment is going, we have transformed and minipreped the truncation mutants (1-193, 1-200) and the point mutant (V175A) and sent those for sequencing. On Friday, we found out that luckily one of samples of ”protein X” 140-215 had the correct sequence so we will be able to maxiprep the bacteria and be ready to use it for future experiments.