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， 已成为发达国家流行病爆发的重大威胁。加剧这些 问题是蚊子种群中杀虫剂抗药性的出现， 在寄生虫种群中，疫苗开发进展迟缓， 蚊子控制计划，以及全球旅行的增加。尖音库蚊复合群中的蚊子 是淋巴丝状线虫寄生虫班氏吴策线虫和几种 虫媒病毒这包括最近在北美引入的西尼罗河病毒。很长的- 这项研究的长期目标是鉴定、分离和描述遗传因素 与滞育相关，滞育是一种强烈影响昆虫地理分布和生存的特征 Culex pipiens complex中的一种。目前，还没有关于分子生物学的知识。 库蚊滞育的决定因素。本建议的目标是基于一般的 假设与滞育表型相关的基因可以通过以下方法鉴定和分离： 定位克隆技术。滞育相关基因的鉴定及认识 从相关生化途径的详细了解， 潜在的应用包括：促进改变的和疾病限制的转基因构建体 种群更替后的行为，有针对性的生物农药的开发，或 鉴定破坏病原体关键行为生物学相关拮抗剂 传播或生存。该项目的具体目标是：1）精细绘制基因组区域 鉴定为含有决定滞育表型的基因，2）鉴定候选基因 决定滞育; 3）对决定滞育的候选基因进行功能分析。 来自遗传杂交的信息将被利用来对抗Cx。致倦鸡基因组 在离散的基因组区域内识别候选基因的项目。候选基因将是 使用微阵列进行评估，然后进行定量真实的时间PCR测定，以确认和扩增 有希望的基因的微阵列结果。将对阳性候选基因进行生物学检测， 相关性使用RNA干扰介导的基因敲低或基因过表达， 重组辛德毕斯转导病毒。实验室实验的结果将是 在代表Cx的自然群体中进行评价。派皮恩斯公司所以Cx。致倦肌

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.