RUI: Molecular and AFM Analysis of a Novel Periplasmic Structure

这项研究的科学价值主要在于提高了对细菌结构的理解。现在很明显，细菌的细胞骨架元素比以前想象的要复杂得多，在过去的10年里，原核生物的这一领域的研究已经产生了重要的信息。一个独特的曲线阵列上的细胞质膜的周质表面上已初步确定的初步研究。 这是可能的，这个网络具有显着的功能，细菌的代谢和生长，它的研究将扩大我们的知识原核processes.A渗透裂解程序已开发的钩虫贪铜菌，结果在生成完整的亚细胞结构，如使用原子力显微镜（AFM）成像。虽然其中一些结构，如细胞幽灵，是已知的，其他的是新的。特别是，已经成像的原生质球样结构在其表面上具有有组织的曲线阵列，表面上由蛋白质组成。曲线阵列由50-250 nm长和30-50 nm宽的短片段组成，这些片段蜿蜒穿过细胞质膜的周质面。曲线阵列本质上可以是动态的，因为当C. necator在基本培养基中生长，它是以短弯曲的片段的形式，但当C. necator生长在丰富的培养基中，初步实验表明它更像是一个网络。该项目的重点是曲线阵列/网络的确认和进一步的结构表征。要做到这一点，优化的条件将被用来获得场发射扫描电子显微镜和透射电子显微镜分析SR原生质球，以确认阵列的存在，并获得更准确的横向尺寸的阵列成分。由于该阵列最近也在大肠杆菌细胞鬼的表面上成像，因此它提出了该阵列是在所有细菌细胞中发现的结构的可能性。 为此，将在不同的细菌物种中进行裂解实验，并采用AFM、SEM和TEM寻找曲线表面网络存在的证据。更广泛的影响这项研究的更广泛的影响是在科学教育领域的学生，一般来说，从弱势教育背景。虽然密歇根州立大学的学生非常有能力，但他们受到不合格的高中教育的阻碍，更重要的是，有限的职业抱负。对于他们中的许多人来说，成为一名研究科学家的想法与成为一名宇航员的机会是同等重要的。参与这个研究项目不仅可以教育他们研究的方式，还可以向他们证明，科学研究的职业生涯并非遥不可及。

Intellectual merit The scientific merit of this research is primarily in an enhanced understanding of bacterial structure. It is now apparent that bacteria are much more complex with regards to cytoskeletal elements than previously thought and this area of prokaryotic research has yielded significant information in the last 10 years. A unique curvilinear array on the periplasmic surface of the cytoplasmic membrane has been, tentatively, identified in preliminary studies. It is possible that this network possesses significant functionalities for bacterial metabolism and growth and its study would expand our knowledge of prokaryotic processes.An osmotic lysis procedure has been developed for Cupriavidus necator that results in the generation of intact subcellular structures, as imaged using atomic force microscopy (AFM). While some of these structures, such as cell ghosts, are known, others are novel. In particular, a spheroplast-like structure has been imaged that possesses an organized curvilinear array on its surface, ostensibly comprised of protein. The curvilinear array is comprised of short 50-250 nm long, and 30-50 nm wide, segments snaking across the periplasmic face of the cytoplasmic membrane. The curvilinear array may be dynamic in nature because when C. necator is grown in minimal medium it is in the form of short curved segments, but when C. necator is grown in rich medium preliminary experiments suggest that it is more of a network. The focus of this project is the confirmation and further structural characterization of the curvilinear array/network. To do this optimized conditions will be used to obtain SR spheroplasts for field emission scanning electron microscopy and transmission electron microscopy analyses to confirm the existence of the array and obtain more accurate lateral dimensions for the array constituents. Because the array has recently also been imaged on the surface of Escherichia coli cell ghosts, it raises the possibility that the array is a structure found in all bacterial cells. To this end, lysis experiments will be conducted in different bacterial species and evidence for the existence of the curvilinear surface network will be sought employing AFM, SEM and TEM. Broader ImpactThe broader impact of this research is in the area of science education of students that are, generally, from disadvantaged educational backgrounds. While the students at MSU are extremely capable, they are hindered by substandard high school educations and, more importantly, limited career aspirations. For many of them the idea of becoming a research scientist is on a par with their chances of becoming an astronaut. Participation in this research project would not only educate them in the ways of research, it would prove to them that a career in scientific research is not unattainable.