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This summer the interns worked on a research project with MSI scientist Pia Abola to develop tools for making vitro biochemical measurements of protein kinase activity. The goals of the project were to (1) create site specific mutations in three of the protein kinases (Sterile7, Kss1 and Fus3) in the alpha mating pheromone pathway MAP kinase cascade, (2) express the mutated proteins in bacteria and, (3) confirm the activity of the mutation by observing the phenotype in yeast cells.

We assigned each student a protein and asked them to generate either a constitutively active or inactive form. The students used site directed mutagenesis and the polymerase chain reaction to generate the mutated proteins. They then used the pET expression system to express the proteins in E.coli bacteria. In the last experiment, they inserted the mutated proteins into yeast cells and assayed the mating response to alpha factor.

At the end of the summer, the students presented their research at a symposium held at the institute [download Powerpoint slides] They also created a wonderful movie chronicling their experiences at Molsci [download/view Quicktime movie].