FIBER DIFFRACTION FROM FILAMENTOUS PLANT VIRUSES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。我们正在收集来自柔性丝状植物病毒的纤维衍射数据，这些数据作为识别，组装，灵活性，病毒-宿主相互作用和方法开发的模型非常宝贵。植物病毒在生物技术方面具有巨大潜力，但开发需要三维结构。纤维衍射通常是唯一可用于确定这种丝状结构的方法。 衍射样品是定向溶胶和干燥纤维。我们发现，为了更好地在干纤维的高度灵活的长丝，并在更刚性的长丝取向溶胶。干燥的纤维需要更少的材料，这是研究低滴度病毒的重要考虑因素。在BioCAT收集宽角和小角衍射数据。 我们获得了菜豆普通花叶病毒和菜豆普通花叶坏死病毒的纤维衍射数据，在样品制备中使用高湿度，在取向和分辨率方面都有显着改善。我们已经获得了更好的模式，从相关的马铃薯Y病毒属大豆花叶病毒（SMV）的两个菌株，这些是第一次从SMV获得的衍射数据，和最好的衍射数据以往任何时候都从重要的马铃薯Y病毒家族的成员。他们将是非常宝贵的，在确定高分辨率马铃薯Y病毒结构。 我们已经从烟草花叶病毒和马铃薯X病毒的定向溶胶中获得了优异的衍射图案，使用新方法制备，所需材料不到传统方法所需材料的5%，并且涉及更少的处理（期刊出版物#3）。该方法将是非常有用的纤维衍射研究时，样品数量有限，不稳定，或危险。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are collecting fiber diffraction data from flexible filamentous plant viruses, invaluable as models for recognition, assembly, flexibility, viral-host interactions, and in methods development. Plant viruses have great potential in biotechnology, but development will require three-dimensional structures. Fiber diffraction is often the only method available for the determination of such filamentous structures. Diffraction samples are oriented sols and dried fibers. We find that order is better in dried fibers of highly flexible filaments, and in oriented sols of more rigid filaments. Dried fibers require much less material, an important consideration for studies of low-titer viruses. Wide and small angle diffraction data are collected at BioCAT. We have obtained fiber diffraction data from bean common mosaic virus and bean common mosaic necrosis virus, significantly improved both in orientation and in resolution by the use of high humidities in specimen preparation. We have obtained even better patterns from two strains of the related potyvirus soybean mosaic virus (SMV); these are the first diffraction data obtained from SMV, and the best diffraction data ever obtained from a member of the important Potyvirus family. They will be invaluable in determining high-resolution potyvirus structures. We have obtained excellent diffraction patterns from oriented sols of tobacco mosaic virus and potato virus X, made using a new method that requires less than 5% of the material needed for conventional methods, and involves much less handling (journal publication #3). The method will be extremely useful in fiber diffraction studies when samples are limited in quantity, labile, or hazardous.