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

智力上的功绩。接合作用包括将DNA直接从供体细胞转移到受体细胞。由于接合元件可以提供自己的遗传物质以及其他驻留的DNA，接合在细菌进化中发挥了深远的作用，使生物体能够获得新的基因，适应新的环境。在接合过程中，DNA通过由接合元件编码的特殊的DNA易位通道在细胞间转移。这个项目将描述保守的ATPase锥，这是枯草杆菌结合元件ICEBs1的DNA转位通道的关键能量成分。该项目旨在揭示哪些结合蛋白与细胞膜相互作用并向细胞膜招募锥体。此外，还将探索连接、三磷酸腺苷水解和定位到膜上所需的锥体序列。这些研究应该会导致发现ICEBs1接合机制的其他关键组件，并更好地理解控制细菌水平基因转移的基本原理。该项目的主要影响是加强本科生的培养，同时拓宽了我们对水平基因转移的理解。这项研究将由本科生在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室课程和高中推广活动中。此外，该项目将允许购买研究级荧光显微镜，这将使学生能够跟踪更广泛的研究兴趣，并学习先进的显微技术。随着更多的学生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，进入生物科学顶级研究生项目和职业生涯的学生比例预计将会增加。

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.