Sex-Specific Gene Activity in the Mosquito Larva

蚊子幼虫的性别特异性基因活性

• 批准号: 6697481
• 负责人: Helen Benes
• 金额: $ 29.04万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6697481
• 关键词: Aedes; DNA footprinting; Drosophilidae; arthropod genetics; binding sites; cell line; fat body; gel mobility shift assay; gender difference; gene expression; genetic mapping; genetic promoter element; genetic regulation; genetic regulatory element; genetic transcription; hemolymph; insect control; larva; reporter genes; secretory protein; tissue /cell culture; transcription factor

DESCRIPTION: (provided by the applicant): The worldwide resurgence of virulent and resistant forms of mosquito-borne diseases has increased the urgency for novel mosquito control strategies. The long-term goal of the proposed research is to elucidate the molecular mechanisms underlying female-specific gene activity in the immature mosquito in order to develop novel mosquito control strategies by transgenic manipulation. Most studies of sex-specific gene activity have focused on expression in adult insects. In the fruit fly, Drosophila melanogaster, the molecular basis for gene activation in adult females and repression in males is well understood for the transcriptional regulator, Doublesex (DSX). Our studies of hexamerin genes have allowed us to develop a model system to study sex-specific gene activity in the larval mosquito. In holometabolous insects, hexamerins (hexameric storage proteins) are abundantly synthesized by the fat body of late-stage larvae. Recently we showed that one of the hexamerins, AatHex-1 .2, produced by fourth-instar larvae of the mosquito, Aedes atropalpus, is truly female-specific. In the 5'-flanking region of the AatHex-1.2 gene, we identified putative regulatory elements (including DSX binding sites), which we believe govern its tissue-, stage- and sex-specific activity. 700 bp of this region are sufficient to direct 10-fold higher reporter gene activity uniquely in the fat body of female larvae of the heterologous dipteran, D. melanogaster. To elucidate the molecular basis for sexually dimorphic AatHex-1.2 activity, we propose to identify the requisite regulatory DNA sequences and transcription factor binding sites. Regulatory sequences will be mapped by gene transfer in the mosquitoes, Aedes aegypti and Ae. atropalpus, and in Drosophila. Transcription factors will be characterized using electrophoretic mobility shift assays and DNase foot printing. Accomplishment of these aims will not only provide a better understanding of the molecular basis for sex- and fat body-specific gene activity in an insect larva but will also define a minimal promoter to improve the sterile insect technique (SIT) for insect control and to develop novel means for driving transgenes into natural populations.

描述：（由申请人提供）：全球范围内致命病毒的死灰复燃 蚊媒疾病的耐药性增加了解决这一问题的紧迫性 新的蚊子控制策略。拟议研究的长期目标 旨在阐明女性特有基因的分子机制 未成熟蚊子的活动，以开发新的蚊子控制方法 通过转基因操作的策略。大多数性别特异性基因的研究 活动集中在成年昆虫中的表达。在果蝇中， 果蝇，成年基因激活的分子基础 女性和男性的压抑对于转录是众所周知的 调节器，双性 (DSX)。 我们对六聚体基因的研究使我们能够开发一个模型系统 研究蚊子幼虫的性别特异性基因活性。在全代谢中 昆虫中，六聚体（六聚体储存蛋白）大量合成 后期幼虫的脂肪体。最近我们展示了其中之一 六聚体，AatHex-1 .2，由蚊子第四龄幼虫产生， 黑须伊蚊确实是女性特有的。在 5' 侧翼区域 AatHex-1.2 基因，我们鉴定了假定的调控元件（包括 DSX 结合位点），我们相信它控制着其组织、阶段和性别特异性 活动。该区域的 700 bp 足以直接提高 10 倍 报告基因活性在雌性幼虫的脂肪体内独特 异源双翅目，D. melanogaster。 为了阐明 AatHex-1.2 两性二态性活性的分子基础，我们 提议鉴定必要的调控 DNA 序列和转录 因子结合位点。调控序列将通过基因转移来绘制 蚊子、埃及伊蚊和伊蚊。萎缩果蝇和果蝇。 将使用电泳迁移率来表征转录因子 位移分析和 DNase 足迹打印。这些目标的实现不会 仅提供对性别和脂肪的分子基础的更好理解 昆虫幼虫体内特定的基因活性，但也将定义最小 促进改进昆虫不育技术（SIT）以控制昆虫和 开发将转基因引入自然群体的新方法。