TRANSFORMATION OF MOSQUITOES WITH PIGGYBACK TRANSPOSON

蚊子传播的传染病继续困扰着人类。这些 疾病对许多人的健康和经济有重大影响 整个世界。控制蚊子媒介的替代战略可以 通过对遗传学和生理学的更多了解而产生的 这些昆虫中。蚊子基因研究的进展将 通过发展一种有效的手段而大大增强 转基因操作蚊子。这其中的一个关键组成部分 能力将是识别昆虫转座子的能力 能够有效地将基因导入蚊子基因组并保持 它处于活动状态。昆虫转座子PiggyBac已经证明 对种类繁多的鳞翅目昆虫进行有效的基因操作 物种。我们还证明了这个转座子能够 显微注射蚊子胚胎中的瞬时运动。这项建议 为一项初步研究寻求短期资金，以探索其潜力 该转座子建立转基因菌株的能力 蚊子物种埃及伊蚊。一个带有 可选择的抗药性基因HSP/OPD已经 被证明对建立转基因昆虫品系有效，将是 通过微量注射将其导入Ae.埃及人的胚胎。.的存在 在G1处选择对氧子后，将检测到转座子 用Southern杂交和PCR扩增的方法， 克隆和测序。关于稳定性和运动性的进一步分析 将对蚊子基因组中的转座子序列进行研究 在时间和进度允许的情况下。这项研究将扩展这一点 实验室探索蚊子转基因的能力，以及 如果成功，将为以下方面提供令人兴奋和急需的能力 显著加强蚊子的基础研究工作 遗传学和生理学。

Mosquito-borne infectious diseases continue to plague mankind. These diseases 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 would be greatly enhanced by the development of an effective means of transgenic manipulation of mosquitoes. A key component of this capability would be the identification of insect transposons that are able to efficiently vector a gene into the mosquito genome and maintain it in an active state. The insect transposon piggyBac has proven effective for genetic manipulation in widely diverse lepidopteran species. We have also shown that this transposon is capable of transient movement in microinjected mosquito embryos. This proposal seeks short-term funding for a pilot study to explore the potential capabilities of this transposon for establishing transgenic strains of the mosquito species, Aedes aegypti. A piggyBac element carrying a selectable insecticide resistance gene, hsp/opd, that has already proven effective for establishing transgenic insect strains, will be introduced by microinjection into Ae. aegypti embryos. The presence of the transposon will be detected following paraoxon selection at the G1 generation using Southern hybridization, and PCR amplification, cloning, and sequencing. Further analyses of the stability and movement of transposon sequences within the mosquito genome will be undertaken as time and progress permits. This research will expand this laboratory's capabilities for exploring transgenesis in mosquitoes, and if successful, will provide an exciting and much needed capability for significant enhancement of the basic research effort in mosquito genetics and physiology.