Post-Winter-Break Plans

What with finals and break, I have had to put my work in the lab on hold for a few weeks; however, this time has given me the opportunity to think about my plans for the future and specifically about what I hope to accomplish next semester.

As mentioned in my last post, our lab recently received a new plasmid with a cDNA cassette encoding the K2 killer toxin, which my previous assays have found to have very strong toxin activity. Since I have had little success transforming yeast strains with K28 plasmids and cassettes, I plan to move forward using this new plasmid. Hopefully, conditions for transformation will be more favorable using a different toxin’s cDNA.

I am also planning to again replicate my competition assays using the toxin-producing strains I currently have, in which the toxins are encoded by cytoplasmic viruses. As mentioned in a previous post, my aim in transforming strains with toxin cDNA cassettes is to yield a system that I can more easily characterize and regulate compared to these virus-encoded systems; however, after so many false starts with my transformations I believe that I should hedge my bets and continue experiments with these toxin-producing strains as well. It may be possible that something about the virus-encoded system that is essential for toxin production is not being replicated by the cDNA cassettes I am trying to use. If I continue work with the virus-encoded systems, I will be able to move forward even if my transformations continue to be unsuccessful.

Finally, over the rest of break I hope to take some time to work on drafting a paper presenting my previous research into biofilm fitness in pure and mixed colonies.