RUI: Characterization of a Conserved ATPase Required for Conjugation of Bacillus subtilis

知识价值。接合涉及DNA直接从供体细胞转移到受体细胞。由于共轭元件可以传递自己的遗传物质以及其他的DNA，因此共轭在细菌进化中发挥了深远的作用，使生物体获得新的基因并适应新的环境生态位。在接合过程中，DNA通过一个由接合元件编码的特殊DNA易位通道从一个细胞转移到另一个细胞。该项目将表征保守的atp酶ConE，这是枯草芽孢杆菌共轭元件ICEBs1 DNA易位通道的关键能量学成分。该项目旨在揭示哪些偶联蛋白与细胞膜相互作用并将ConE招募到细胞膜上。此外，将探索缀合，ATP水解和定位到膜所需的ConE序列。这些研究将导致发现ICEBs1偶联机制的其他关键组成部分，并更好地理解控制细菌水平基因转移的基本原理。更广泛的影响。本项目的主要影响是加强本科生的培养，同时拓宽我们对水平基因转移的理解。该研究将由本科生在PI的指导下设计、实施和分析。PI将继续将探究性研究融入她的本科生物化学实验课程和高中拓展活动中。此外，该项目将允许购买一个研究级荧光显微镜，这将使学生遵循更广泛的研究兴趣和学习复杂的显微镜技术。进入顶级研究生课程和从事生物科学职业的学生比例预计会增加，因为更多的学生将有机会获得有用的研究技能，展示他们的工作，并体验科学发现的兴奋。

Intellectual Merit. Conjugation involves the transfer of DNA directly from a donor cell to a recipient cell. Since conjugative elements can deliver their own genetic material as well as other resident DNA, conjugation has played a profound role in bacterial evolution, allowing organisms to acquire new genes and adapt to new environmental niches. During conjugation, DNA is transferred from cell to cell through a specialized DNA translocation channel that is encoded by the conjugative element. This project will characterize the conserved ATPase ConE, a critical energetics component of the DNA translocation channel of the conjugative element ICEBs1 of Bacillus subtilis. This project is designed to reveal which conjugation proteins interact with and recruit ConE to the cell membrane. Furthermore, the sequences of ConE required for conjugation, ATP hydrolysis, and localization to the membrane will be explored. These studies should lead to the discovery of other critical components of the ICEBs1 conjugation machinery and a better understanding of the fundamental principles that govern horizontal gene transfer in bacteria.Broader Impacts. Major impacts of this project are to enhance undergraduate student training while broadening our understanding of horizontal gene transfer. The research will be designed, conducted, and analyzed by undergraduate students with guidance from the PI. The PI will continue to integrate inquiry-based research into her undergraduate biochemistry laboratory course and high school outreach activities. Furthermore, this project will permit the purchase of a research-grade fluorescence microscope which will allow students to follow a wider range of research interests and learn sophisticated microscopy techniques. The percentage of students entering top-tier graduate programs and careers in the biosciences is expected to increase, as many more students will have the opportunity to gain useful research skills, present their work, and experience the excitement of scientific discovery.