FIBER DIFFRACTION FROM FILAMENTOUS PLANT VIRUSES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们正在收集柔性丝状植物病毒的纤维衍射数据，作为识别、组装、灵活性、病毒与宿主相互作用的模型以及方法开发的宝贵模型。植物病毒在生物技术方面有很大的潜力，但发展需要三维结构。纤维衍射通常是确定这种丝状结构的唯一方法。 衍射样品为取向的溶胶和干燥的纤维。我们发现，在高弹性长丝的干燥纤维中，以及在较坚硬的长丝的取向溶胶中，有序性较好。干燥的纤维需要的材料要少得多，这是研究低滴度病毒的重要考虑因素。在BioCAT收集了广角和小角衍射数据。 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，通过在样品制备中使用高湿度，显著地提高了取向度和分辨率。我们从两个相关的马铃薯Y病毒大豆花叶病毒(SMV)株系中获得了更好的图谱；这是首次从SMV获得的衍射数据，也是有史以来从重要的马铃薯Y病毒家族成员获得的最好的衍射数据。它们在确定高分辨率马铃薯Y病毒结构方面将是无价的。 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极好的衍射图，该方法使用了一种新的方法，所需材料不到传统方法所需的5%，并且涉及的处理要少得多。当样品数量有限、不稳定或危险时，该方法将在纤维衍射研究中非常有用。

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 . The method will be extremely useful in fiber diffraction studies when samples are limited in quantity, labile, or hazardous.