ASGSB 2003 Annual Meeting Abstracts


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SURFACE BASED CHARACTERIZATION OF GENE TRANSCRIPTION USING ACOUSTIC WAVE NETWORK ANALYSIS.  C.N. Jayarajah and M. Thompson, Department of Chemistry, University of Toronto, Canada.

   It is important to develop surface biochemical techniques to monitor gene transcription in merging nano and biotechnologies in space biology.  We have developed an on-line acoustic wave biosensor method to probe gene transcription using the T7 bacteriophage model system.  In this method, the promoter DNA is immobilized on the gold electrode surface coating of a piezoelectric quartz crystal.  The binding of the RNA polymerase and subsequent mRNA synthesis is monitored by fitting the impedance data from the crystal to an equivalent circuit model. From this fitting, the resonance frequency and motional resistance of the biochemical processes coupled to the quartz crystal are extracted and used to analyze the biochemical interactions involved.  We have also characterized the surface based synthesis of mRNA using radiolabelling and gel electrophoresis.  In addition, cyclic voltammetry and scanning probe microscopy have been employed to characterize the surface features towards a better understanding of the acoustic wave results.  We demonstrate that the acoustic wave biosensor is a viable real time, label-free method to efficiently characterize DNA promoter RNA polymerase interactions and the synthesis of mRNA in gene transcription.  As well, this sensor has been used to study drug binding to DNA in inhibiting transcription.  The real time network analysis data allows for the calculation of on and off rates for these binding interactions.  These studies will be relevant to developing analytical tools for gene chips; as well as multiplexing, automation, and miniaturization towards building efficient biotechnological tools for space life sciences research.

(Supported by NSERC Canada)

 

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