Klee Quantifies Proteins at the University of Florida

Posted: February 24, 2016

Penn State Ph.D. student Sara Klee describes her six-week study at the University of Florida Cancer and Genetics Research Center.
Photo credit: Sara Klee

Photo credit: Sara Klee

A Graduate Student Enhancement Grant allowed me to spend six weeks in November and December at the University of Florida (UF) Cancer and Genetics Research Center in Gainesville learning to quantify proteins using a method called iTRAQ.  I am currently characterizing a protein in Erwinia amylovora, the fire blight pathogen, which has not been studied in any plant pathogen and likely alters the expression of other proteins.

To understand what proteins are differentially present in my mutant bacteria compared to wild type, I was invited to Sixue Chen’s lab to learn this advanced proteomics technique.  We were able to both identify and quantify most of the proteins present in our bacteria.  This is a major advantage over looking at protein expression levels individually, which would be very expensive and time-consuming.  I worked under his visiting scientist Islam Mostafa, learning to label and fractionate my isolated proteins as well as analyze this complex set of data.  In total, we identified over 2,000 proteins, many of which were differentially expressed in the mutant.

I also took advantage of my time at UF by attending seminars in the Department of Biology and learning unrelated techniques, such as viral-induced gene silencing.   At the Chen lab’s meetings I learned how proteomics are being applied beyond the scope of my own project, such as how plants acclimate to saline stress.