FUNCTIONS OF THE PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS

人呼吸道合胞病毒蛋白质的功能

• 批准号: 6288863
• 负责人: PETER LEON COLLINS
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6288863
• 关键词: DNA directed RNA polymerase; RNA biosynthesis; genetic transcription; microorganism culture; microorganism growth; nucleocapsid; phosphoproteins; plasmids; posttranslational modifications; protein structure function; replicase; respiratory syncytial virus; tissue /cell culture; transfection; virus RNA; virus envelope; virus genetics; virus protein; virus replication

Human respiratory syncytial virus (RSV) is an enveloped RNA-containing virus of the paramyxovirus family of Order Mononegavirales (the nonsegmented negative strand RNA viruses). RSV is the most important viral agent of pediatric respiratory tract disease worldwide. Other members of the Order include measles, mumps, rabies and Ebola viruses. The RSV genome is a single negative strand of RNA of 15,222 nucleotides that encodes 10 major mRNAs and 11 proteins, as we have shown previously. The purpose of this project is to identify the functions of the viral proteins and to reconstruct events in the viral growth cycle under conditions where they can be more readily studied. Knowledge of the functions of the proteins is important for a basic understanding of this important human pathogen and for the design of live-attenuated recombinant vaccine viruses.We previously developed an experimental system for RSV based on the intracellular coexpression and assembly of viral RNA and protein components from recombinant plasmid vectors. This involves transfecting cultured cells with plasmids which individually encode a truncated version of negative-sense genomic or positive-sense antigenomic RNA (the latter being an intermediate in RNA replication) as well as whatever mix of RSV proteins is desired. In this way, various steps in the viral replication cycle can be reconstituted, such as genome replication, gene expression, gene regulation, packaging and infection. Because each component is expressed individually, each can be manipulated individually with regard to relative amount as well as by mutagenesis for structure-function studies. - virus, respiratory tract disease, infectious disease, pediatrics, vaccines, live- attenuated viral vaccine, recombinant DNA, proteins

人呼吸道合胞病毒（RSV）是一种含有Mononegavirales的帕托马伏病毒家族的包膜含RNA的病毒（未分割的阴性链RNA病毒）。 RSV是全球小儿呼吸道疾病的最重要的病毒剂。该命令的其他成员包括麻疹，腮腺炎，狂犬病和埃博拉病毒。 RSV基因组是15,222个核苷酸的RNA的单个负链，它编码10个主要mRNA和11种蛋白质，如我们先前所示。该项目的目的是确定病毒蛋白的功能，并在可以更容易研究的条件下重建病毒生长周期中的事件。对蛋白质功能的了解对于对这种重要人类病原体的基本理解以及对活衰减的重组疫苗病毒的设计很重要。我们先前根据基于细胞内的共表达和来自重组质粒源的病毒RNA和蛋白质成分的细胞内共表达和蛋白质成分的RSV实验系统。这涉及用质粒转染培养的细胞，这些质粒分别编码负义基因组或阳性抗原组RNA的截短版本（后者是RNA复制中的中间体）以及RSV蛋白的任何混合。这样，可以重新组建病毒复制周期中的各个步骤，例如基因组复制，基因表达，基因调节，包装和感染。因为每个组件都是单独表达的，所以每个组件都可以单独操纵相对量以及通过诱变进行结构功能研究。 - 病毒，呼吸道疾病，传染病，儿科，疫苗，活衰减病毒疫苗，重组DNA，蛋白质