BUNYAVIRUS/MOSQUITO INTERACTIONS

描述(改编自申请人的摘要)：本报告的总体目标 研究是检查最初的分子相互作用，这是一个关键 确定本雅氏病毒感染蚊子能力的第一步 向量。布尼亚病毒是人类疾病的一个重要原因 世界各地的家畜。大多数布尼亚病毒都会传播给人类 动物被蚊子叮咬。他们将使用La Crosse脑炎病毒(LACV)和 其天然蚊媒三色伊蚊用于这些研究。他们会 研究病毒糖蛋白在病毒与蚊子结合中的作用 中肠，在艾氏的感染中。Triseriatus的LACV，并在专一性 对选定蚊子物种的LACV。它们会完全产生传染性的LACV 使用最近使用的基于RNA聚合酶1的系统从克隆的cdna中提取 对流感病毒进行反向工程。他们将引入特定的突变 和嵌合片段和基因进入LACV。基于大量的工作 在他们的实验室里，他们假设La Crosse的G2包膜蛋白 病毒是病毒与蚊子中肠细胞结合的关键，并且 因此对蚊子感染至关重要。他们将确定蛋白质序列 与La Crosse病毒的中肠细胞结合所需的G2基因 它的天然蚊子媒介，以及另一种允许的蚊子物种。他们 将确定载体的特异性是否在一定程度上是由多肽介导的 病毒糖蛋白中的序列，如果这些序列与 确定病毒与合格媒介蚊子中肠细胞的结合。这个 高效生产布尼亚病毒的技术研究进展 特定的突变将使个体基因的体内研究成为可能 在蚊子感染和哺乳动物发病中的作用

DESCRIPTION (Adapted from the Applicant's Abstract): The overall goal of this research is to examine the initial molecular interactions that are a critical first step in determining the ability of a bunyavirus to infect ist mosquito vector. Bunyaviruses are a significant cause of disease in humans and in domestic animals worldwide. Most bunyaviruses are transmitted to humans and animals by mosquitoes. They will use La Crosse encephalitis virus (LACV) and its natural mosquito vector Aedes triseriatus for these studies. They will examine the role viral glycoproteins play in the binding of virus to mosquito midgut, in the infection of Ae. triseriatus by LACV, and in the specificity of LACV for selected mosquito species. They will generate infectious LACV entirely from cloned cDNA using a RNA polymerase 1-based system that was recently used to reverse engineer influenza viruses. They will introduce specific mutatuions and chimeric segments and genes into LACV. Based on a significant body of work in their laboratory, they hypothesize that the G2 envelope protein of La Crosse virus is critical for virus binding to the cells of the mosquito midgut, and thus critical for mosquito infection. They will determine the protein sequences of G2 that are required for biding of La Crosse virus to the midgut cells of its natural mosquito vector, and to another permissive mosquito species. They will determine if vector specificity, in part, is mediated by pepetide sequences in the viral glycoproteins and if these sequences are the same that determine virus binding to midgut cells of competent vector mosquitoes. The development of efficient techniques to generate bunyaviruses containing specific mutations will permit in vivo investigations of individual gene functions in mosquito infections and in mammalian pathogenesis