Continuing Biofilm/Toxin Assays: Progress!

I have been putting a lot of time into lab so far this semester. A lot of my work has been towards generation of my Trojan Horse strain. I have done transformations with both an extrachromosomal plasmid and with a cassette targeted for integration into the yeast genome. I am now assaying potentially successful transformants for toxin activity. As well, I have processed copies of my current K2-cassette plasmid, in which the cassette is under an inducible promoter, to generate a new plasmid in which the toxin cassette should be under a constitutive promoter. I next need to have the resulting cassette sequenced to check its composition. If successful, I should be able to transform yeast strains to express K2 either constitutively or when induced, allowing me to perform a number of different experiments using the K2 toxin to disrupt biofilms.

I am also continuing work on assays with the K2- and K28-producing strains I currently have. I successfully grew colonies with these strains, alone and in combination with biofilm-producing strains, and took plugs to store colonies for quantification of final strain numbers. Because we were concerned that continuing toxin activity would affect the numbers for mixed colonies in storage, I had to process all of these colonies immediately after taking plugs. This resulted in a very long late-night lab session (over 16 hours to take plugs and process mixed colonies)!

I still have a lot of lab work to do towards my research goals. I have also started writing for the Materials and Methods sections of both my Honors thesis and a paper which Dr. Murphy and I plan to submit for publication, and started finding and organizing reference papers to cite in the introductory and background sections of my thesis.