Transgenic engineering of Aedine mosquitoes

伊丁蚊的转基因工程

• 批准号: 6738965
• 负责人: Malcolm J. FRASER
• 金额: $ 40.95万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6738965
• 关键词: Aedes; arthropod genetics; biotechnology; genetic manipulation; genetically modified animals; transfection /expression vector; transposon /insertion element

DESCRIPTION (provided by applicant): Mosquito-borne infectious diseases continue to have significant impact on the health and economy of much of the world. Alternative strategies for control of mosquito vectors could arise through a greater understanding of the genetics and physiology of these insects. Progress in genetic research with mosquitoes will be greatly enhanced by the development of effective and reproducible means of transgenic manipulations. A key component of this capability is the identification and manipulation of insect transposons that are able to efficiently vector genes into the mosquito genome, permit gene tagging, and maintain genes in an active state. The convergence of research in this field has resulted in the successful transgenic manipulation of mosquitoes with several insect transposons, each with its own unique properties. The insect transposon piggyBac has been demonstrated to be effective for genetic manipulation in widely diverse insect species, including mosquitoes. Recently, we demonstrated transformation of Aedes acgypti with piggyBac using the cinnabar eye coloration marker gene. This proposal seeks significant funding to understand and exploit the capabilities of the piggyBac transposon for establishing and maintaining transgenic strains of Aedine mosquito species. Additional piggyBac-based transgene vectors containing desirable genes under the control of either ubiquitous insect promoters or tissue-specific mosquito promoters will be constructed, tested, and combined with the selectable markers for transgenesis. The presence and expression of the transposon and vectored genes will be detected following selection for the co-transforming marker gene at the Gi generation, and analyzed using a variety of techniques including Southern hybridization, PCR amplification and sequencing, Northern and Western analyses, and immunoflourescence, as appropriate. Genetic characterization of the stability and movement of transgene sequences within the mosquito genome will be examined over several generations. This research will provide an exciting and much needed capability for significant enhancement of the basic research effort in mosquito genetics and physiology.

描述（由申请人提供）：蚊子传染病 继续对许多大多数人的健康和经济产生重大影响 世界。可能会出现控制蚊子向量的替代策略 通过对这些的遗传学和生理学有更深入的了解 昆虫。通过蚊子的遗传研究进展将大大增强 通过开发有效且可重复的转基因手段 操纵。此功能的关键组成部分是标识和 操纵能够有效矢量基因的昆虫转座子 进入蚊子基因组，允许基因标记并维持活跃的基因 状态。该领域的研究的融合导致了成功 用几个昆虫转座子对蚊子的转基因操纵，每个蚊子 具有自己独特的属性。昆虫转座子piggybac一直是 事实证明对广泛多样的昆虫的基因操纵有效 物种，包括蚊子。最近，我们证明了 使用cinnabar Eye Marker基因的Acgypti与Piggybac一起使用。这 提案寻求大量资金来理解和利用能力 Piggybac Transposon的建立和维持转基因菌株 亚丁蚊子的物种。基于Piggybac的其他转基因向量 在两种无处不在的昆虫的控制下包含理想的基因 将构建，测试启动子或组织特异性蚊子启动子 并与可选的转基因标记结合在一起。存在和 遵循转座子和矢量的基因的表达 选择GI代的共转化标记基因，并选择 使用包括南部杂交，PCR在内的多种技术进行分析 放大和测序，北部和西方分析以及 适当的免疫荧光。稳定的遗传特征 将检查蚊子基因组中的转基因序列的运动 超过几代。这项研究将为令人兴奋而多 需要大量提高基础研究工作的必要能力 蚊子遗传学和生理学。