From the last post I mentioned I am glad to report I have substantial data which helps shed light on the entire purpose of my research. First of all, I have completed my Opp mutant growth trail and viability assay. From the results I have collected, I can conclude that glutathione (GSH) cannot enter within the cytoplasm of Opp mutants in great enough concentrations to cause any stress to the bacteria. This is seen as 0,4 and 8 mM GSH all had the same growth and viability. This confirms that Opp gene that encodes of the oligopeptide permease transporter is not only crucial but necessary for GSH stress.
I have also made a significant discovery in my assessment of GSH concentration in the cytoplasm of my model bacterium BCG. Using this fluorescent kit I was able to quantify approximately the amount of GSH that was entering the cytoplasm of the bacteria at 4 and 8 mM using 0 mM as a control. What I found was that the mount of GSH entering the cytoplasm at 4 mM was not statistically different than 0MM. However 8mM GSH we observed 5.73 fold increase in GSH content . These results help confirm the hypothesis that our bacteria, BCG is experiencing reductive stress from GSH and is unable to handle such stress when the concentration is 8mM and higher.
This project has greatly increased my understanding of the scientific process. After much failure and restarting experiments, I have learned that science does not always follow a straight path. And in order to become a successful scientist you need to have patience and respect for the scientific process.