RESPIRATORY SYNCYTIAL VIRUS POLYPEPTIDE SYNTHESIS

呼吸道合胞病毒多肽合成

• 批准号: 3134543
• 负责人: DENNIS M LAMBERT
• 金额: $ 11.5万
• 依托单位: JAMES N. GAMBLE INSTITUTE OF MEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-01-01 至 1986-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3134543
• 关键词: Pneumovirus; chemical structure function; gel electrophoresis; microorganism immunology; peptide chemical synthesis; phosphorylation; virus RNA; virus envelope; virus infection mechanism; virus replication

This proposal will be an investigation of respiratory syncytial (RS) virus polypeptide synthesis. The kinetics of viral polypeptides synthesized in cells infected at high multiplicity (5-10 PFU/cell) will be followed by pulse-labeling and pulse-chase experiments. Acetone precipitated and immunoprecipitated viral polypeptides will be analyzed on polyacrylamide slab gels. Comparisons of virion structural polypeptides with in vivo and in vitro translation products will be made and correlation of infected cell proteins with cell-free products will be carried out by peptide mapping and immunoprecipitation with monospecific antibodies. Intracellular nucleocapsids will be analyzed so that proteins associated with viral RNA throughout the infectious cycle can be identified and compared with the proteins of virion nucleocapsids. Phosphorylationof viral proteins will be studied in infected cells and in purified virions. Experiments designed to isolate and purify virion glycoproteins will be carried out so that monospecific antisera can be made. Monospecific antibodies will be used to determine the biological functions of the glycoproteins in the infectious process. The information gained from these experiments may (1) benefit the development of viral or subviral component vaccines as well as (2) improve diagnostic techniques. Immunological and electrophoretic techniques will be utilized to attempt to determine differences in the polypeptides of various strains of RS virus.

本研究将对呼吸道合胞病毒（RS）进行研究 多肽合成 病毒多肽合成的动力学 以高多重性（5-10 PFU/细胞）感染的细胞将随后进行 脉冲标记和脉冲追踪实验。 丙酮沉淀， 将在聚丙烯酰胺上分析免疫沉淀的病毒多肽 平板凝胶。 病毒体结构多肽与体内和体外的比较 将制备体外翻译产物，并将感染细胞 具有无细胞产物的蛋白质将通过肽图谱进行， 用单特异性抗体进行免疫沉淀。 细胞内 将对核衣壳进行分析，以便与病毒RNA相关的蛋白质 在整个感染周期中，可以识别并与 病毒粒子核衣壳蛋白质。 病毒蛋白的磷酸化 在感染的细胞和纯化的病毒体中进行了研究。 实验旨在 分离和纯化病毒体糖蛋白将被进行， 可以制备单特异性抗血清。 单特异性抗体将用于 确定糖蛋白的生物学功能， 过程 从这些实验中获得的信息可能（1）有利于 病毒或亚病毒组分疫苗的开发以及（2）改善 诊断技术。 免疫学和电泳技术将 用于试图确定多肽的差异， 各种RS病毒株。