My current research projects focus on elucidating structure and function of molecules of interest using molecular biological techniques. One project involves the cloning and sequencing of the gene for GAPDH (Glyceraldehyde-3- Phosphate Dehydrogenase) from several common vegetable plants. The GAPDH gene codes for an enzyme in the Glycolytic Pathway and is present not only in plants, but also animals, fungi, bacteria, and protists, and the gene that codes for it is highly conserved. Our lab has cloned and sequenced the GAPDH gene from kohlrabi, a member of the cabbage family. Genomic DNA was isolated from plant leaves and amplified using PCR. The purified gene was then cloned into the pJet plasmid (Bio Rad) and subsequently sequenced. The GAPDH genes from yellow squash and yellow bush beans are also currently under study in our lab. We have also cloned and parially sequenced the GAPDH gene from marigold and we are currently attempting the same with the diving dolphin plant.
Another current project is using site-directed mutagenesis to study of structure-stability relationships in proteins. Our lab uses the model protein, ribonuclease T1, for these studies and our focus has been on the sole alpha helix in the structure.