ISAT/Geog Study Abroad In Malta 2006
2006 Field Study Projects
TITLE
Defensin Mutagenesis in Malta
ABSTRACT
The Defensin Mutagenesis project begins with a Polymerase Chain Reaction (PCR) on isolated plasmid DNA. The first part of this project consisted of finding the optimal concentration of MgCl2 found in the buffer to be used later, and the optimal annealing temperature, that will give the most permissive reaction. Forward and reverse primers were created for three different binding sites for the desired exons. Three MgCl2 and annealing temperature conditions were tested and matched with the three exons' binding sites for a total of nine reactions. PCR was performed on these nine reactions and the results were run on an agarose gel. The results of electrophoresis predicts the optimal MgCl2 and annealing temperature combination to be used later in the experiment. In the second part of the project, point mutations were preformed on the defensin DNA. PCR was performed and the results were then run on an agarose gel, and gel purification was performed to remove the desired gene. A parent plasmid was digested with restriction enzymes. The gene were ligated into the parent plasmid and transformed into competent cells (E.Coli). The clones were plated, incubated, and selected for DNA purification and sequencing.
TEAM
Amy Babiarz, ISAT '08
Jennifer McCool, ISAT '07
SPONSORS/ADVISORS
Dr. Pierre Schembri-Wismayer, Department of Anatomy and Cell Biology, University of Malta
Dr. George Coffman, Assistant Professor, ISAT, JMU
Dr. Bob Kolvoord, Professor, ISAT, JMU
FINAL PRESENTATION (PDF)
