STUDIES WITH SYNTHETIC NEURAMINIDASE SUBSTRATES

合成神经氨酸酶底物的研究

• 批准号: 2059824
• 负责人: Peter Palese
• 金额: $ 21.61万
• 依托单位: MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-05-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2059824
• 关键词: MDCK cell; enzyme mechanism; enzyme substrate; exo alpha sialidase; fatty acylation; genetic library; genetic manipulation; host organism interaction; human genetic material tag; influenzavirus A; laboratory mouse; laboratory rabbit; method development; microorganism hemagglutinin; molecular cloning; mutant; palmitates; protein structure function; site directed mutagenesis; transfection; virulence; virus genetics; virus protein; virus replication

The development of a reverse genetics system for influenza viruses allows us to site-specifically mutate the genomes of these negative strand RNA viruses. Thus, we are now able to perform structure-function studies on proteins in the context of infectious virus and to define specific virulence characteristics on a molecular level. Using mutational analysis, we will study the functional and antigenic determinants of the influenza virus neuraminidase (NA) in the cytoplasmic tail, the stalk and the head region. The characterization of genetically manipulated influenza viruses (transfectants) will define in a more precise way how the NA influences host range or neurovirulence. Similar studies will be done on the hemagglutinin (HA) in order to define the role of palmitoylation of the HA in the formation of infectious particles and to study host range and virulence as it is influenced by the HA. In order to facilitate the rapid isolation of transfectants with changes in non- HA/non-NA genes, attempts will be made to develop convenient systems which will allow for the rapid exchange of any influenza virus gene with a cDNA-derived RNA segment. We will also develop protocols to establish a reverse genetics system for influenza B viruses.

用于流感病毒的反向遗传系统的开发允许 我们要特异性地突变这些负链RNA的基因组 病毒。因此，我们现在能够对 蛋白质在传染病毒的背景下，定义特定 分子水平上的毒力特征。使用突变 分析，我们将研究的功能和抗原决定因素 细胞质尾巴，茎和 头部区域。遗传操纵的表征 流感病毒（转染物）将以更精确的方式定义 NA会影响宿主范围或神经电动机。类似的研究将是 在血凝素（HA）上完成 HA的棕榈酰化在传染性颗粒的形成中 研究寄主范围和毒力受HA的影响。为了 为了促进转染剂的快速分离，随着非 - HA/非NA基因，将尝试开发便捷的系统 这将允许任何流感病毒基因的快速交换 cDNA来源的RNA段。我们还将开发建立协议 流感病毒的反向遗传系统。