In vitro transcription and transcription factors binding studies via single molecule analysis

ANYL 145

Hua Yu1, Tian Wu2, Daniel Forrest, Rod Runnheim, Chris Churas, Karl E. Hauschild3, Aseem Ansari3, and David C. Schwartz4. (1) Laboratory for Molecular and Computational Genomics, Department of Chemistry, Biotechnology Center, University of Wisconsin - Madison, 425 Henry Mall, Madison, WI 53706, (2) School of Pharmacy, University of Wisconsin - Madison, Madison, WI 53705, (3) Department of Biochemistry, University of Wisconsin - Madison, Madison, WI 53706, (4) Laboratory for Molecular and Computational Genomics , Department of Chemistry, Laboratory of Genetics, UW Biotechnology, University of Wisconsin - Madison, 425 Henry Mall, Madison, WI 53706
Transchip is an in vitro transcription system designed to track detailed biochemical action occurring on a whole genome basis using single molecule templates derived from genomic DNA. High-throughput fluorescence microscopy combined with tools originally developed for Optical Mapping (OM) are being used to image and quantitate DNA-protein interactions and nascent transcriptional complexes on ensembles of single molecule templates bound to surfaces. New surface modalities were developed to enable biochemical actions such as transcription on a surface platform. Image analysis was done after restriction enzyme digestion to find the loci of transcriptional complexes (reflective of promoters) on a variety of DNA templates. Characterization of single DNA molecules bearing transcriptional complexes was done via RNase digestion, fluorescent-labeling of nascent RNA and immuno-labeling to the RNA Polymerase directly on surfaces. AFM analysis revealed detail structural information of individual transcriptional complexes. Transcription processes that involve transcription factors (Ubx-Exd) binding to DNA were also studied based on single molecule FRET experiments.
 

General Session
7:00 PM-9:00 PM, Sunday, 26 March 2006 Georgia World Congress Center -- Ex. Hall B4, Poster

Sci-Mix
8:00 PM-10:00 PM, Monday, 27 March 2006 Georgia World Congress Center -- Ex. Hall B4, Sci-Mix

Division of Analytical Chemistry

The 231st ACS National Meeting, Atlanta, GA, March 26-30, 2006