CAREER:Translocation and Unwinding by DnaB

在这个CAREER提案中，PI建议使用DNA活性的单分子操作测量来研究DNA复制的机制。该提案的具体科学目标是：1。确定dna易位的机制。2. 确定dsDNA解绕的机制。（PI将通过施加不稳定的机械力来改变dsDNA的磨损平衡，并测量由此产生的解卷速度来研究这个问题。）3. 开发一种新的单分子技术，允许多个单分子测量并行执行。该研究结果将为未来研究整个复制复合体的机制行为奠定基础，并将与理解高等生物的复制动力学相关。该提案的教育目标是通过引入从物理科学到生物探针的本科生和研究生，培养下一代跨学科，以生命科学为导向的研究人员。这将通过结合本提案研究目标的实验室实习和课堂教学来完成。PI将培训研究生和本科生，他们将学习生物学、生物化学和物理学的技术，以便进行实验。参与的本科生将从不同的背景中招募，包括那些在科学领域代表性不足的学生。

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.