SINDBIS VIRUS DETERMINANTS OF INFECTION IN MOSQUITOS

描述（改编自申请人的摘要）：申请人提出 研究辛德毕斯病毒与埃及伊蚊的相互作用。为此他们 将使用经过改造的SIN分子克隆来表达 表型特征，并携带方便的标记基因。 他们将检查SIN的两个感染性DNA克隆，其中一个感染伊蚊 中肠无效（2 J病毒）和另一种感染中肠 并在蚊子中引起传播性感染（MRE 1001 病毒）。MRE 1001病毒是2 J病毒的非结构基因之间的嵌合体， 病毒和来自中肠感染菌株MRE 16的结构基因 株2 J菌株和嵌合MRE 1001菌株均已被鉴定。 经工程改造以含有第二亚基因组RNA，该第二亚基因组RNA与正常衣壳3-引物 编码GFP（绿色荧光蛋白）的区域 口服或注射给药时这三种病毒的生长曲线 路线，并比较器官向性，外源性潜伏期， 病毒的传播潜力。他们将创造更多的 MRE 1001和2 J菌株的结构基因的嵌合体，逐基因 以确定中肠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 一个相关的研究方向，他们建议创建一个新的感染性DNA克隆， 来自MRE 16菌株的SIN。该菌株与MRE 1001菌株的不同之处在于 所有的非结构基因和结构基因 中肠复制传播型病毒株

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.