STUDIES WITH SYNTHETIC NEURAMINIDASE SUBSTRATES

流感病毒反向遗传学系统的发展， 我们对这些负链RNA的基因组进行位点特异性突变， 病毒因此，我们现在能够进行结构-功能研究， 在感染性病毒的背景下， 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簇的 流感病毒神经氨酸酶（NA）在细胞质尾部，茎和 头部区域。基因操纵的表征 流感病毒（转染子）将以更精确的方式定义 NA影响宿主范围或神经毒力。类似的研究将 对血凝素（HA）进行研究，以确定 在感染性颗粒的形成中HA的棕榈酰化， 研究宿主范围和毒力，因为它受到HA的影响。为了 为了便于快速分离具有非- HA/非NA基因，将尝试开发方便的系统 这将允许任何流感病毒基因与 cDNA衍生的RNA片段。我们还将制定协议， 一种针对B型流感病毒的反向遗传学系统。

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.