J.P. Manalo: Protein Identification and DNA Replication
J.P. Manalo —This summer, I am continuing an independent research project (begun in the spring semester) in the Chemistry Department at Wabash on the role of proteins in biomineralization events in plants.
Under the guidance of Dr. Aaron Wyman, I’ve been able to isolate proteins that associate with grape raphides, which are biominerals composed of calcium oxalate and produced naturally in grapes.†Calcium oxalate is also a major component of human kidney stones. Thus, the prime focus of this research is to investigate potential mechanisms to alleviate the pain caused by kidney stones by identifying proteins that control the growth of calcium crystals.
Over the summer, I have been working to identify the optimal conditions for amplifying the grape genes corresponding to numerous isolated raphide associating proteins (RAPs) using PCR, in an effort to clone them.PCR stands for polymerase chain reaction, and it is a method that allows scientists to replicate small amounts of DNA accurately and in large quantities.
The resulting PCR products are then analyzed using agarose gel electrophoresis in order to determine their size and number of base pairs.†From this work, we next intend to sequence these DNA fragments and continue to clone full-length copies of these genes in an effort to determine if their expressed protein products influence grape raphide size and morphology.
Working with Dr. Wyman and another summer student researcher, Ahson Ali, has been a great experience for me, as our research with biominerals is set in a pleasant and light environment.†The number of laboratory techniques that I’ve learned this summer will be beneficial to me in my future endeavors when I plan to pursue a career in the medical field. The coursework and lab work at Wabash College definitely prepared me for this summer experience.
I would like to thank the Chemistry Department at Wabash College for granting me this opportunity to perform research this summer. I would especially like to thank Dr. Wyman for selecting me to do research with him and for mentoring me over the summer.