Monday, April 14, 2008

08 Science Fair: 1st Place for Biotechnology

First Place: Erin Kim, 11th Grade
“Engineering of Stayphylococcus aureus Sortase A to improve protein detection and localization” (13005)
Category: Biochemistry
Region: Mathematics and Science High School, Clover Hill, Richmond

Abstract: Proteins serve as catalysts that maintain metabolic processes in the cell. Because of its important roles, new methods to characterize protein localization and trafficking are required to better understand the many complex regulatory pathways. Current technologies rely heavily upon Green Fluorescent Protein (GFP), a bulky fluorescent protein with a propensity to polymerize within a cell, and a chemical coloring method which chemically dyes the proteins. However, these two primary methods have problems: GFP is limited in its utility because of its size, which interferes with proteins’ regular movement or pathways, and the chemical coloring method lacks specificity because it labels all proteins. In this experiment, the researcher alternatively combined a fluorescent substrate with Staphylococcus aureus Sortase A (Srt A) to create a better tool for protein detection and localization. The results showed that Srt A with fluorescent substrate could be used as a protein marker. However, in vitro studies using the preferred fluorescent substrate have verified limited use of Srt A. To increase Srt A’s efficiency, it was put into successful rounds of mutagenesis. Error prone PCR was used to create random mutagenesis and it will allow the researcher to screen for active mutated enzyme with the researcher’s desired specificity and properties. The result from the experiment will provide researchers with a better tool in which to study any protein of interest when tagged with the specificity marker.

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