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）是单负链病毒目（Mononegavirales）副粘病毒科的一种含包膜RNA的病毒（非节段负链RNA病毒）。呼吸道合胞病毒（RSV）是世界范围内小儿呼吸道疾病最重要的病毒因子。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的单负链，编码10种主要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质的功能，并在更容易研究的条件下重建病毒生长周期中的事件。的蛋白质的功能的知识是重要的基本了解这一重要的人类病原体和减毒活重组疫苗virus.We的设计以前开发了一个实验系统RSV的细胞内共表达和组装的基础上的病毒RNA和蛋白质组分从重组质粒载体。这涉及用质粒转染培养的细胞，所述质粒单独编码截短形式的反义基因组RNA或正义反基因组RNA（后者是RNA复制的中间体）以及所需的任何RSV蛋白质混合物。通过这种方式，可以重建病毒复制周期中的各个步骤，例如基因组复制、基因表达、基因调控、包装和感染。因为每种组分都是单独表达的，所以每种组分都可以在相对量方面单独操作，也可以通过诱变进行结构-功能研究。- 病毒，呼吸道疾病，感染性疾病，儿科，疫苗，减毒活疫苗，重组DNA，蛋白质