FUNCTIONS OF THE PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS

人呼吸道合胞病毒（RSV）是单病毒目副粘病毒科（无节段负链RNA病毒）中一种含包膜RNA的病毒。RSV是全球儿童呼吸道疾病最重要的病毒病原体。该教团的其他成员包括麻疹、腮腺炎、狂犬病和埃博拉病毒。RSV基因组是一条15222个核苷酸的单负链RNA，编码10个主要mrna和11个蛋白质，如我们之前所示。该项目的目的是确定病毒蛋白的功能，并在更容易研究的条件下重建病毒生长周期中的事件。了解蛋白质的功能对于基本了解这一重要的人类病原体和设计减毒重组活疫苗病毒非常重要。我们之前基于重组质粒载体的病毒RNA和蛋白质组分在细胞内的共表达和组装开发了RSV的实验系统。这包括用质粒转染培养细胞，质粒分别编码负义基因组RNA或正义抗基因组RNA的截断版本（后者是RNA复制的中间物）以及所需的RSV蛋白混合物。通过这种方式，可以重构病毒复制周期的各个步骤，如基因组复制、基因表达、基因调控、包装和感染。因为每个成分都是单独表达的，所以每个成分都可以根据相对量进行单独操作，也可以通过诱变进行结构-功能研究。-病毒、呼吸道疾病、传染病、儿科、疫苗、减毒活疫苗、重组DNA、蛋白质

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