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.
Figure 1. Crystal structure of Apo-Glyceraldehyde-3-Phosphate
Dehydrogenasefrom Palinurus Versicolor; PDB 1CRW.
Figure 2. Amplified GAPDH genes: Lane 1 = Control; Lane 2 = PCR
product from Kohlrabi before cleaning; Lane 3 = PCR from Kohlrabi after
purification; Lane 4 = 500 base pair ladder.
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.
Figure 3. Restriction Digest of pJet plasmid with Kohlrabi GAPDH gene insert.
Lanes 1 and 4, 500 base pair ladder; Lane 2, undigested plasmid DNA; Lane 3,
digested plasmid DNA – higher band is the pJet plasmid and the lower band is
the GAPDH gene insert.