Poster abstracts

Poster number 18 submitted by Katharine Stahon

Kinetic Analysis of T7 RNA Polymerase from Novel DNA Templates

Katharine E. Stahon (Chemistry, John Carroll University), Stephanie Clack (Chemistry, John Carroll University), Michael P. Martin (Biology, John Carroll University), David P. Mascotti (Chemistry, John Carroll University)

Abstract:
Typical in vitro transcription assays use templates that are made of linear, double-stranded DNA. In this study we used novel templates that possessed a double-stranded promoter for T7 RNA polymerase recruitment, but the remainder of each DNA construct contained three different downstream variations. The control template utilized a linear, completely double-stranded DNA template. This was compared to two templates of the same length where one template strand was self-complementary and formed a hairpin (snapback), whereas the other strand possessed no self-complementary and remained single-stranded. All three templates produced RNA that was similar in length by denaturing polyacrylamide gel analysis. The kinetic data for each template could be fit by relatively simple Michaelis-Menten parameters. We found variation among the templates utilized at different temperatures. Results will be shown.

Keywords: T7 RNA Polymerase, kinetics