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）：全球有毒的复兴 蚊子传播疾病的抗性形式增加了 新颖的蚊子控制策略。拟议研究的长期目标 是阐明女性特异性基因的分子机制 未成熟蚊子的活性，以发展新型蚊子控制 通过转基因操纵的策略。性别特异性基因的大多数研究 活性集中在成年昆虫的表达上。在果蝇中， 果蝇Melanogaster，成人基因激活的分子基础 雄性的女性和抑制作用已被很好地理解 调节器Doublesex（DSX）。 我们对己酰胺基因的研究使我们能够开发一个模型系统 在幼虫蚊子中研究性别特异性基因活性。在金代谢中 昆虫，六聚蛋白（六聚体储存蛋白）大量合成 后期幼虫的脂肪体。最近我们表明了其中一个 六胺，AATHEX-1 .2，由蚊子的第四instar幼虫生产 艾德斯attropalpus确实是特定于女性的。在5'乘区域 AATHEX-1.2基因，我们确定了推定的调节元素（包括DSX 绑定位点），我们认为它控制其组织，阶段和性别特定的组织 活动。该区域的700 bp足以指导高10倍 记者基因活性在雌性幼虫的脂肪体中唯一 异源Dipteran，D。Melanogaster。 为了阐明性二态AATHEX-1.2活性的分子基础，我们 建议识别必要的调节DNA序列和转录 因子结合位点。调节序列将通过基因转移映射 蚊子，埃及埃及和AE。阿托帕尔普斯和果蝇。 转录因子将使用电泳迁移率来表征 换档测定和DNase脚印。实现这些目标不会 仅提供对性别和脂肪的分子基础的更好理解 昆虫幼虫中的身体特异性基因活性，但也将定义最小 启动子改善无菌昆虫技术（SIT），以进行昆虫控制和 开发新颖的手段将转基因驱动到自然种群中。