SINDBIS VIRUS DETERMINANTS OF INFECTION IN MOSQUITOS

辛毕斯病毒蚊子感染的决定因素

• 批准号: 6028112
• 负责人: KENNETH E OLSON
• 金额: $ 20.69万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6028112
• 关键词: Aedes; Sindbis virus; clone cells; communicable disease transmission; disease vectors; gene expression; genetic strain; green fluorescent proteins; host organism interaction; polymerase chain reaction; site directed mutagenesis; structural genes; transfection /expression vector; virus genetics; virus infection mechanism; virus receptors; virus replication

DESCRIPTION (Adapted from the Applicant's Abstract): The applicants propose to study the interactions of Sindbis virus with Aedes aegypti. To do this they will use molecular clones of SIN that have been engineered to express phenotypic characteristics of interest and which bear convenient marker genes. They will examine two infectious DNA clones of SIN, one of which infects Aedes midguts inefficiently (the 2J virus) and the other which infects midguts efficiently and causes disseminated infections in the mosquito (the MRE1001 virus). The MRE1001 virus is a chimera between he nonstructural genes of the 2J virus and the structural genes from a midgut infecting strain, the MRE16 strain. The 2J strain and the chimeric MRE1001 strains have both been engineered to contain a second sub-genomic RNA 3-prime to the normal capsid region which encodes GFP (green fluorescent protein) They will first compare growth curves of these three viruses when given by the oral or injection routes, and compare the organ tropisms, extrinsic incubation periods, and transmission potentials of the viruses. They will then create additional chimeras, gene by gene, of the structural genes of the MRE1001 and 2J strains to define the genetic determinants of midgut tropism and dissemination. Finally, they will introduce site specific mutations and deletions to further refine the genetic determinants of viral gut replication and dissemination. In a related line of research they propose to create a new infectious DNA clone of SIN from the MRE16 strain. This strain will differ from the MRE1001 strain in that all the non-structural genes as well as the structural genes will derive from a mid-gut replicating, disseminating virus strain.

描述（改编自申请人的摘要）：申请人提议 研究辛德毕斯病毒与埃及伊蚊的相互作用。为了做到这一点，他们 将使用经过工程改造的 SIN 分子克隆来表达 感兴趣的表型特征并带有方便的标记基因。 他们将检查 SIN 的两个传染性 DNA 克隆，其中之一感染伊蚊 中肠效率低下（2J 病毒），另一种感染中肠 有效并导致蚊子传播感染（MRE1001 病毒）。 MRE1001病毒是2J非结构基因之间的嵌合体 病毒和中肠感染株 MRE16 的结构基因 拉紧。 2J菌株和嵌合MRE1001菌株均已被 被设计为含有正常衣壳的第二个亚基因组 RNA 3 引物 编码 GFP（绿色荧光蛋白）的区域 他们将首先比较 这三种病毒口服或注射时的生长曲线 途径，并比较器官趋向性、外源性潜伏期和 病毒的传播潜力。然后他们将创建额外的 MRE1001 和 2J 菌株结构基因的嵌合体，逐个基因 定义中肠趋向性和传播的遗传决定因素。 最后，他们将引入位点特异性突变和缺失以进一步 完善病毒肠道复制和传播的遗传决定因素。在 他们提出的一项相关研究旨在创建一种新的传染性 DNA 克隆 来自 MRE16 菌株的 SIN。该菌株与 MRE1001 菌株的不同之处在于 所有非结构基因和结构基因都将派生 来自中肠复制、传播的病毒株。