TRANSFORMATION OF MOSQUITOES WITH PIGGYBACK TRANSPOSON

蚊媒传染病继续困扰人类。这些 疾病对许多国家的健康和经济产生重大影响， 世界控制蚊媒的替代战略可 通过对遗传学和生理学的更好理解而产生 这些昆虫。蚊子基因研究的进展将 通过发展有效的方法， 蚊子的转基因操作。其中一个关键组成部分是 能力将是鉴定昆虫转座子， 能够有效地将基因导入蚊子基因组， 它处于活跃状态。昆虫转座子piggyBac已经证明 对广泛多样鳞翅目昆虫有效的遗传操作 物种我们还表明，这种转座子能够 显微注射蚊子胚胎的短暂运动。这项建议 寻求短期资金进行试点研究，以探索 这种转座子建立转基因株系的能力 埃及伊蚊一个piggyBac元件， 选择性杀虫剂抗性基因hsp/opd， 被证明是建立转基因昆虫品系的有效方法， 通过显微注射引入Ae.埃及胚胎的存在 转座子将在G1对氧子选择后被检测到 使用Southern杂交和PCR扩增产生， 克隆和测序。对稳定性和运动性的进一步分析 蚊子基因组内的转座子序列的研究将在 只要时间和进度允许。这项研究将扩大这一 实验室探索蚊子转基因的能力， 如果成功，将提供一个令人兴奋的和急需的能力， 大大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.