Genetics of Dengue Vector Competence in Aedes aegypti

描述(申请人提供)：登革热是最迅速的 热带地区疾病不断扩大，超过20亿人面临风险。 登革热病毒(DEN)的传播涉及到 主要蚊媒埃及伊蚊、病毒及其人类的基因组 主持人。拟议的研究将侧重于其中的两个组成部分，即 研究Ae.媒介能力的遗传学。埃及伊蚊和程度 载体的能力受DEN病毒的遗传变异的影响。我们 定位了影响Ae能力的数量性状基因座(QTL)。宙斯盾 中肠感染登革2型病毒(DEN2)。这两种肠道感染 屏障(MIB)QTL被称为dmib2、dmib3，被定位在染色体II和 三、分别是。拟议工作的主要目标是使用 标记辅助选择(MAS)产生4株Ae。埃格普里。这些 将是一株在两个座位上都易感的纯合子菌株(Ds2)，一株纯合子菌株 两个座位均难治(DR2)和2个纯合子在一个座位上易感 另一位点(DMIB2、DMIB3)为纯合子耐火材料。这些菌株将 提供了一个极好的机会来阐明与 在DEN传递基础上的QTL上的每个基因型。DS2将用于 从世界范围内的Ae群体中分离和定位洞穴屏障QTL。退伍军人至 确定额外的QTL是否控制DEN敏感性和/或是否替代 等位基因存在于目前发现的QTL上。DMIB2、DMIB3和DR2将 用来评估两个基因座上的等位基因是如何单独和共同赋予 4种不同DEN血清型和不同基因型别的敏感性 血清型。DS2和DR2将用于构建重组自交系 用于消减杂交法推测克隆cDNA的品系 由dmib3编码，并协助协作进行定位克隆 Dmib3.

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.