RUI: Characterization of Metabolic Genes Involved in Bacillus Sporulation

在饥饿期间，一些细菌物种经历了一种细胞分化，导致一种高度弹性和休眠的形式，称为孢子。由于多种原因，一些细菌的孢子是有害的，因此对孢子形成的研究很重要。该项目的主要目标是发现饥饿的枯草芽孢杆菌细胞如何仍然能够获得能量来为孢子形成提供燃料。核心假设是，当产孢枯草芽孢杆菌细胞消耗其储存的葡萄糖时，母细胞代谢基因（mmg）集群编码的酶通过消化细胞自身的脂肪酸来提供能量和物质。由mmg集群指定的酶将被分离出来，它们参与脂肪酸消化的必要步骤的能力将被检查。为了确定这些基因对产孢的影响，将测量缺乏这些基因的产孢突变体的能量状态。对这些突变体进行基因阵列实验，还将发现冗余的或支持mmg簇的产孢基因。这项研究的成功完成将具有重要意义，因为这一知识将有助于对产孢过程中使用的其他代谢途径进行新的研究。更广泛的影响：该项目公布的结果将为以前未研究的基因提供缺失的化学数据，从而有助于提高公共枯草芽孢杆菌基因组序列数据库的实用性。这些数据还将提高整套基因组数据库的预测能力，这反过来将影响依赖基因组学的广泛的生物技术领域。也许这个项目对科学和社会最重要的影响将来自于将在北卡罗来纳大学格林斯博罗分校接受该项目培训的大量不同的化学和生物化学学生。当他们最终开始自己的科学事业时，学生参与者将通过贡献他们将从这个项目中学到的广泛的沟通和科学技能来造福科学和社会。这个跨学科项目将通过鼓励学生在工业领域从事科学事业或在nsf资助的领域进行研究生水平的研究来促进发现，同时促进教学和学习。这个实验室以前的学生现在正在攻读有机化学和生物工程的博士学位，在生物技术公司工作，或者在高中教科学。新生将继续完成所有的实验工作，使用不同的技术，包括细菌培养、DNA和蛋白质方法、分析化学和合成有机化学。学生还将获得与职业生活、研究生学习和科学诚信相关的强有力的职业指导。首席研究员是第一代大学毕业生，将继续招募和指导不同群体的本科生和硕士生，其中将包括代表性不足的群体和第一代大学生。例如，这个大型研究小组迄今为止由62%的女性和46%的代表性不足的少数民族组成。学生将通过在科学文献中广泛传播他们的研究成果和在科学会议上的演讲，获得书面和口头交流的技能。该项目还将加强UNCG的教育和研究基础设施，不仅通过学生的研究机会，而且通过生物化学讲座和实验室课程。这些课程将使用这项研究来教授代谢途径以及用于研究它们的方法。最后，这项研究将通过为公共资助的枯草芽孢杆菌基因组数据库中可用的理论基因功能分配添加所需的实验数据来造福社会。

During periods of starvation some bacterial species undergo a type of cell-differentiation which results in a highly resilient and dormant form called a spore. The study of sporulation is important since the spores of several bacterial species are harmful for a wide range of reasons. The main objective of this project is to discover how starving Bacillus subtilis cells still manage to have the energy to fuel sporulation. The central hypothesis is that when sporulating B. subtilis cells deplete their stores of glucose, the enzymes encoded by the mother cell metabolic gene (mmg) cluster contribute energy and material through the digestion of the cell's own fatty acids. The enzymes specified by the mmg cluster will be isolated, and their ability to participate in the necessary steps of fatty acid digestion will be examined. To determine the effects these genes have on sporulation, the energy state of sporulating mutants which lack these genes will be measured. Sporulation genes which are redundant or function in support of the mmg cluster will also be discovered by gene array experiments with these mutants. Successful completion of this research will be significant, because this knowledge will enable new investigations of other metabolic pathways used during sporulation.Broader Impact: Published results from this project will provide missing chemical data for previously unstudied genes, and will thus help improve the utility of the public genome sequence database for B. subtilis. Such data will also improve the predictive power of the overall set of genome databases, which will in turn impact the broad array of biotechnological fields which depend on genomics. Perhaps the most significant impact of this project on science and society will come from the large and diverse group of chemistry and biochemistry students that will receive training from this project at UNC-Greensboro. As they ultimately embark on their own science careers, the student participants will benefit science and society by contributing the broad range of communication and scientific skills they will learn from this project. The interdisciplinary project will advance discovery while promoting teaching and learning by inspiring students to pursue science careers in industry or graduate level research in NSF-sponsored fields. Former students from this lab are currently pursuing Ph.D's in organic chemistry and biological engineering, are working at biotechnology companies, or teaching high school science. New students will continue to perform all of the lab work, using diverse techniques including bacterial culture, DNA and protein methods, analytical chemistry, and synthetic organic chemistry. The students will also receive strong career mentoring related to professional life, graduate school, and scientific integrity. The Principal Investigator, a first-generation college graduate, will continue to recruit and mentor a diverse group of undergraduate and Masters students, which will include underrepresented groups and first-generation university students. For example, this large research group has been comprised so far of 62% female and 46% underrepresented minorities. The students will gain skills in written and oral communication through wide dissemination of their research in the scientific literature and presentations at scientific meetings. The project will also enhance the educational and research infrastructure at UNCG, not only from the student research opportunities, but also through the biochemistry lecture and laboratory courses. These courses will use this research to teach metabolic pathways along with the methods used to study them. Finally, this research will benefit society by adding needed experimental data to the theoretical gene-function assignments available in the publicly-funded B. subtilis genome database.