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Capillary Electrophoresis Identification of Rhodium-DNA Interstrand Cross-linkage

Alec Brisbois, Dr. Shari Dunham and Dr. Alison Holliday, Department of Biochemistry, Moravian College, 1200 Main Street Bethlehem, PA 18018

The treatment of cancer and the development of novel drugs to eradicate this collection of abnormal cells continues to baffle researchers to this day. One potential drug, a rhodium complex, inhibits cell division by interacting with DNA. This interaction could be interstrand or intrastrand binding; in order to determine the amount of interstrand binding that occurs, methods such as slab gel electrophoresis are used to separate metal modified DNA. However, the tagging/staining methods often used to visualize these metal-DNA adducts either require a radioactivity license or employ fluorescence that may be quenched by the rhodium. In order to visualize Rh-modified DNA, another form of gel electrophoresis known as capillary gel electrophoresis (CGE) is promising, as this method uses UV light absorbance to visualize the compounds instead of a fluorescent stain/tag. This research project seeks to determine if CGE is able to separate rhodium modified DNA from unmodified DNA, whether these results are comparable to standard gel electrophoresis, and if the percentage of interstrand Rh-DNA adducts can be measured. Preliminary findings have shown that single strand DNA of different lengths can be resolved at the single nucleotide level using CGE. However, thus far, our data suggest that duplex DNA has denatured only partly in CGE. In the analysis of the Rh-modified DNA, separate peaks are expected for unmodified denatured DNA, denatured Rh-modified DNA with interstrand linkages, and denatured Rh-modified DNA with intrastrand linkages. The percent of bound rhodium will be calculated in order to determine the effectiveness of the Rh compound as a DNA binding agent, and eventually, an anticancer drug. 

 

 




Additional Abstract Information

Presenter: Alec Brisbois

Institution: Moravian College

Type: Poster

Subject: Biochemistry

Status: Approved


Time and Location

Session: Poster 2
Date/Time: Mon 3:00pm-4:00pm
Session Number: 2507