The 20th annual Symposium will be held on Thursday, April 23, 2009.

Fluorescence Analysis of the Twort Group 1 Introns using an ABI Prism 310 Genetic Sequencer
Poster Session-Science Complex Atrium, 4:00 PM

Tim  Stevens,   '10 88
   Majors: Biochemistry, Biology
   Hometown: Grosse Pointe Park, MI

Sponsor(s): Christopher Rohlman
Support: FURSCA

Abstract: 
One of the dogmas of modern biology is that deoxyribonucleic acid (DNA) and ribonucleic acids (RNA) code for genetic information. DNA can be expressed through transcription, the process of turning DNA into RNA. RNA can be processed by excising sequences called introns, resulting in a finished molecular product that can function in the cell. Introns are linear sequences of ribonucleic acid (RNA) that are spliced out after their biological transcription and before the RNA is used in our cells. Group I introns are catalytic RNAs capable of performing a range of phosphotransesterification reactions including self-splicing and RNA cleavage. Biological catalysts are essential to life because the conditions in our body would make many essential reactions impossible in the absence of the catalyst. Kinetic data is recorded over varying reaction conditions to better understand the catalytic nature of the Twort group I intron, and data is acquired by combining several techniques. These techniques include capillary electrophoresis, fluorescence, and laser spectroscopy. The kinetic data in my research is recorded with the use of an ABI 310 genetic sequencer. The sequencer depends on capillary electrophoresis to separate DNA or RNA molecules based on size. This allows the bound ribozyme, unbound ribozyme, and the substrate to separate due to variations in their size. To quantify the binding results, a laser in the sequencer emits a light of appropriate wavelength to be a photon donor to excite the fluorescently labeled substrate. The excited substrate quickly returns to its normal state with the emission of a photon. The wavelength and strength of this emission is recorded by the sequencer. This produces quantifiable data that allows the binding behavior of the Twort group I intron to be better understood by establishing a model of catalysis. Catalytic models for reactions in living organisms are essential to comprehending the biochemistry that makes life possible.

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