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（根据申请人的摘要改编）：申请人建议 研究信德比斯病毒与伊德斯伊德岛的相互作用。为此，他们 将使用已设计的罪分子克隆来表达 感兴趣的表型特征和具有方便的标记基因。 他们将检查两个传染性的DNA罪恶克隆，其中一个感染了艾德斯 中肠效率低下（2J病毒），另一种感染了中肠病 有效地引起蚊子的传播感染（MRE1001 病毒）。 MRE1001病毒是2J的非结构基因之间的嵌合体 病毒和来自中肠感染菌株的结构基因MRE16 拉紧。 2J菌株和嵌合MRE1001菌株均为 设计为在正常的衣壳上包含第二个亚基因组RNA 3-PRIME 编码GFP（绿色荧光蛋白）的区域将首先比较 口服或注射给予这三种病毒的生长曲线 路线，并比较器官的偏向主义，外部孵化期和 病毒的传输电位。然后他们将创建其他 MRE1001和2J菌株的结构基因的基因的嵌合体，基因 定义中肠tropism和传播的遗传决定因素。 最后，他们将介绍特定地点的突变和删除以进一步 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线，旨在创建一个新的感染性DNA克隆 MRE16菌株的罪。该菌株将与MRE1001菌株不同 所有非结构基因以及结构基因都将得出 从中期复制，传播病毒菌株。