CAREER:Translocation and Unwinding by DnaB

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In this CAREER proposal the PI proposes to study the mechanisms of DnaB replication using single molecule manipulation measurements of the DNA activity. The specific scientific aims of the proposal are: 1. Determine the mechanism of DnaB translocation. 2. Determine the mechanism of dsDNA unwinding. (The PI will study this question by altering the fraying equilibrium of dsDNA through application of a destabilizing mechanical force, and measuring the resulting unwinding velocity.) 3. Develop a novel single-molecule technique that will allow multiple single-molecule measurements to be performed in parallel. The results of the proposed research will lay the groundwork for future studies of the mechanistic behavior of the entire replication complex and will be relevant for understanding replication dynamics in higher organisms. The educational aim of the proposal is to train the next generation of interdisciplinary, life science oriented researchers by introducing undergraduate and graduate students from the physical sciences to biological probes. This will be accomplished through both laboratory internships integrated with the research aims of this proposal, and through classroom teaching. The PI will train both graduate and undergraduate students, who will learn techniques from biology, biochemistry and physics in order to carry out the experiments. Participating undergraduate students will be recruited from several backgrounds, including those that are underrepresented in the sciences.