HOMOLOGOUS GENE TRANSFER SYSTEMS FOR THE MOSQUITO

这项工作将有助于了解生殖 蚊子是人类和动物的重要媒介， 疾病，包括疟疾、黄热病、登革热、脑炎和 其他世界各地。 重组DNA技术的出现 这让人们对蚊子的基因改造产生了希望， 目的是防止致病性病毒的传播 有机体 因此，我们必须了解基因 在这种昆虫中受到调节。 这项工作将侧重于控制 在分子水平上的基因表达。 很长 研究的长期目标是为发展 这项技术将允许对蚊子进行基因操作。 在蚊子繁殖中起作用的基因提供了一种 发展分子生物学和遗传学的基本工具 这种重要的害虫。 本建议的重点是 在卵发育过程中表达的雌性特异性基因， 对类固醇激素20-羟基蜕皮激素的反应 我们计划 在我们已经确定的早期研究的基础上， 基因组文库，4个克隆编码卵黄蛋白原基因-这是 仅在脂肪体中表达，以响应蜕皮激素， 另一个克隆，含有蜕皮激素诱导基因， 尤其是在卵泡细胞中。 该提案的一个重要目标是描述5'- 这些基因的调控序列，最终目标是 开发可用于定义核苷酸区域的构建体， 赋予蜕皮激素反应性和基因组织特异性 表情 由于基因表达的功能分析， 蚊子需要有效的转化技术， 其发展受到严重阻碍， 克隆蚊子的基因，这项建议的第二个主要目标是， 优化蚊子转染系统， 的表达。 很明显，蚊子的基因改造是一种 考虑到提供的强大工具，目标是合理的 重组DNA技术，以及 各种各样的植物和动物的转变。 给定 蚊子在医学上的重要性， 昆虫作为遗传转化特定靶标， 很明显

This work will contribute to an understanding of the reproductive physiology of the mosquito, an important vector of human and animal diseases including malaria, yellow fever, dengue, encephalitis and others worldwide. The advent of recombinant DNA technology has raised the hope that the mosquito could be genetically engineered, with the aim of preventing transmission of disease-causing organisms. It is, therefore, vital that we understand how genes are regulated in this insect. This work will focus on the control of gene expression at the molecular level in a mosquito. A long term goal of the research is to contribute to the development of technology that will allow the genetic manipulation of mosquitoes. Genes that play a role in mosquito reproduction provide an essential tool for developing the molecular biology and genetics of this important pest insect. The present proposal is focused on female-specific genes that are expressed during egg development in the response to the steroid hormone, 20-hydroxyecdysone. We plan to build upon earlier studies in which we have identified, from genomic libraries, 4 clones encoding vitellogenin genes - which are expressed only in the fat body in response to ecdysone - and another clone, containing an ecdysone-inducible gene expressed specifically in ovarian follicle cells. An important goal of this proposal is to characterize the 5'- regulatory sequences of these genes, with the ultimate goal of developing constructs that can be used to define nucleotide regions that confer ecdysone responsiveness and tissue-specificity of gene expression. Since functional analysis of gene expression in mosquitoes requires efficient transformation techniques, the development of which has been severely hampered by the lack of cloned mosquito genes, a second major goal of this proposal is to optimize a mosquito transfection system to allow the rapid analysis of hormone-inducible gene constructs in vitro. It is clear that genetic transformation of mosquitoes is a reasonable goal given the powerful tools made available by recombinant DNA technology, and the rapid advances being made in the transformation of a wide variety of plants and animals. Given the obvious medical importance of the mosquito, the choice of this insect as a particular target for genetic transformation is apparent.