Investigation of the Nodavirus lifecycle in vivo

诺达病毒体内生命周期的研究

• 批准号: 7052795
• 负责人: Jason Kenneth Lanman
• 金额: $ 4.88万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7052795
• 关键词: RNA virus; computer simulation; electron microscopy; fluorescence microscopy; life cycle; postdoctoral investigator; protein structure; protein structure function; three dimensional imaging /topography; tomography; virus genetics; virus infection mechanism; virus protein

DESCRIPTION (provided by applicant): Flock House Virus (FHV) is a non-enveloped, icosahedral RNA virus that infects insects. FHV has been thoroughly studied genetically, biophysically, and structurally, which makes it an excellent system for extending these studies to investigate the virus lifecycle (virus entry, disassembly, assembly, and cellular exit) in situ. To study the FHV lifecycle in situ the virus coat protein containing the tetracysteine tag will be labeled specifically with biarsenical fluorophores, which are visible in both the fluorescent and electron microscopes allowing the different images to be correlated. Additionally, in optical pulse-chase experiments the FHV will be successively labeled with different colored biarsenical fluorophores to distinguish older and younger coat protein. To investigate at higher resolution the spatial relationship of FHV within cells, tomographic electron microscope images will be used to produce three-dimensional reconstructions of the virus infected cells.

描述（由申请人提供）： Flock House病毒（FHV）是一种未发育的二十面体RNA病毒，可感染昆虫。 FHV已在遗传，生物物理和结构上进行了彻底研究，这使其成为扩展这些研究以原位研究病毒生命周期（病毒进入，拆卸，组装和细胞出口）的绝佳系统。为了研究FHV生命周期的原位，将特异性标记包含四环素星态标签的病毒涂层蛋白与Biarsenical荧光团标记，在荧光和电子显微镜中可见，允许不同的图像相关。此外，在光学脉冲追踪实验中，FHV将依次用不同的彩色biarsenical荧光团标记，以区分较旧的和年轻的外套蛋白。为了在较高的分辨率下研究细胞内FHV的空间关系，层析成像电子显微镜图像将用于产生病毒感染细胞的三维重建。