Diapause in Culex Mosquitoes

项目摘要 蚊子传播的疾病是大多数发展中国家的主要健康问题，已经 成为发达国家疫情暴发的重大威胁。加剧了这些问题 问题是蚊子种群中出现了抗药性、抗药性 在寄生虫种群中，疫苗开发进展缓慢，缺乏对 蚊子控制计划，以及增加全球旅行。库蚊复合体中的蚊子 是淋巴丝状线虫寄生虫Wuchereria bancrofti和几个 虫媒病毒。这包括最近在北美引入的西尼罗河病毒。长的- 拟议研究的长期目标是识别、分离和表征遗传因素。 与滞育有关，这一特征强烈影响了白纹伊蚊的地理分布和存活率 淡色库蚊复合体内的种类。目前，还没有关于分子的知识。 库蚊滞育的决定因素。这项建议的目标是基于一般的 与滞育表型相关的基因可以通过以下方式识别和分离的假设 位置克隆技术。决定滞育的基因鉴定及其知识 根据对相关生化途径的详细了解，可能有几个 潜在的应用包括：促进改变和疾病限制的转基因结构 人口置换后的行为，开发有针对性的生物杀虫剂，或 干扰病原菌关键行为的生物相关拮抗剂的鉴定 传播还是生存。该项目的具体目标是：1)精细绘制基因组区域图 确定含有决定滞育表型的基因，2)确定候选基因 3)对决定滞育的候选基因进行功能分析。 来自基因杂交的信息将被用来对抗CX。西洋参基因组 确定离散基因组区域内候选基因的项目。候选基因将是 使用微阵列进行评估，然后进行实时定量聚合酶链式反应分析以确认和扩展 微阵列结果显示前景看好的基因。阳性候选基因将被检测为生物学 使用RNA干扰介导的基因敲除或基因过度表达的相关性 重组辛德比斯转导病毒。实验室实验的结果将会是 在代表CX的自然种群中进行评估。罗布麻属(Piniens S.S.)和CX.[医]五倍子属

Project Summary Mosquito-borne diseases represent major health problems in most developing countries and have emerged as significant threats for epidemic outbreaks in developed countries. Exacerbating these problems are the emergence of insecticide resistance in mosquito populations, drug resistance among parasite populations, delayed progress in vaccine development, lack of support for mosquito control programs, and increased global travel. Mosquitoes in the Culex pipiens complex are primary vectors for the lymphatic filarioid nematode parasite, Wuchereria bancrofti and several arboviruses. This includes the recently introduced West Nile virus in North America. The long- term objectives of the proposed research are to identify, isolate and characterize genetic factors associated with diapause, a trait that strongly impacts the geographic distribution and survival of species within the Culex pipiens complex. At present, no knowledge exists on the molecular determinants of diapause in Culex. The objectives of this proposal are based on the general hypothesis that genes associated with the diapause phenotype can be identified and isolated by positional cloning techniques. Identification of genes determining diapause and the knowledge from the detailed understanding of the associated biochemical pathways could have several potential applications including: transgene constructs that promote altered and disease limiting behaviors following population replacement, development of targeted biopesticides, or identification of biologically relevant antagonists that disrupt behaviors critical to pathogen transmission or survival. The specific aims of the project are to: 1) fine-scale map genome regions identified as containing genes that determine the diapause phenotype, 2) identify candidate genes determining diapause, and 3) conduct functional analysis of candidate genes determining diapause. Information from genetic crosses will be leveraged against the Cx. quinquefasciatus genome project to identify candidate genes within discrete genome regions. Candidate genes will be evaluated using microarrays, followed by quantitative real time PCR assays to confirm and expand microarray results for promising genes. Positive candidate genes will be assayed for biological relevance using RNA interference mediated gene knockdowns or gene overexpression using recombinant Sindbis transducing viruses. Results from laboratory experiments will then be evaluated among natural populations representing Cx. pipiens s.s. and Cx. quinquefasciatus.