Genetics of Dengue Vector Competence in Aedes aegypti

描述（由申请人提供）：登革热是一种最迅速的 热带地区疾病的蔓延使超过20亿人处于危险之中。 登革病毒（DEN）的传播涉及病毒与宿主之间的复杂相互作用。 主要蚊子载体埃及伊蚊、病毒及其人类的基因组 主持人拟议的研究将侧重于其中两个组成部分， 研究了Ae.埃及和程度， 所述载体能力受DEN病毒中遗传变异的影响。我们 已经定位了影响Ae.阿吉普里 中肠感染登革2型病毒（DEN 2）。这两种中肠感染 屏障（MIB）QTL被称为dmib 2、dmib 3，并被定位到染色体II和 三是分别。拟议工作的主要目标是使用 分子标记辅助选择（MAS）筛选出4个Ae. Aegypri。这些 将是在两个基因座（DS 2）上易感的纯合菌株， 两个基因座（DR 2）均为难治型，一个基因座为纯合易感型 另一个基因座为纯合子难治型（DMIB 2、DMIB 3）。这些菌株将 提供了一个特殊的机会来阐明与 每个基因型的QTL基础登革传播。DS 2将用于 从世界各地山羊草群体中分离和定位DEN屏障QTL。阿吉普里岛 确定额外的QTL是否控制DEN易感性和/或是否替代 等位基因存在于当前鉴定的QTL处。DMIB 2、DMIB 3和DR 2将 用于评估两个基因座上的等位基因分别和一起如何赋予 在不同的DEN血清型和基因型的敏感性，在每个4 血清型DS 2和DR 2将用于构建重组近交系 用于消减杂交方法以克隆纯化的cDNA的细胞系 编码的bydmib 3，并协助协作努力， DMIB 3。

DESCRIPTION (provided by applicant): Dengue fever is one of the most rapidly expanding diseases in the tropics with over 2 billion people at risk. Transmission of Dengue virus (DEN) involves a complex interaction between the genomes of the primary mosquito vector, Aedes aegypti, the virus, and its human host. The proposed research will focus on two of these components by examiningthe genetics of vector competence in Ae. aegypti and the degree to which vector competence is affected by genetic variance in the DEN virus. We have mapped Quantitative Trait Loci (QTL) affecting the ability of Ae. aegypri midgut to become infected with Dengue-2 virus (DEN2). These twomidgut infection barrier (MIB) QTL are called dmib2, dmib3 and were mapped to chromosomes II and III respectively. The primary goal of the proposed work is to use marker-assisted selection (MAS) to generate 4 strains of Ae. aegypri. These will be a strain homozygous susceptible at both loci (DS2), a strain homozygous refractory at both loci (DR2) and 2 strains homozygous susceptible at one locus and homozygous refractory at the other locus (DMIB2, DMIB3). These strains will provide an exceptional opportunity to elucidate the phenotypes associated with each genotype at the QTL underlying DEN transmission. DS2 will be used to isolate and map DEN barrier QTL from worldwide populations of Ae. aegypri to determine if additional QTL control DEN susceptibility and/or if alternate alleles exist at the currently identified QTL. DMIB2, DMIB3, and DR2 will be used to assess how alleles at both loci separately and together confer susceptibility in different DEN serotypes and genotypes in each of the 4 serotypes. DS2 and DR2 will be used in the construction of recombinant inbred lines for use in a subtractive hybridization approach to clone cDNAs putatively encoded bydmib3and to assist in a collaborative effort to positionally clone dmib3.