HOMOLOGOUS GENE TRANSFER SYSTEMS FOR THE MOSQUITO

这项工作将有助于理解生殖 蚊子的生理学--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 世界各地的其他人。重组DNA技术的出现 带来了这种蚊子可以通过基因工程改造的希望， 目的是防止致病传播 有机体。因此，至关重要的是我们要了解基因是如何 在这种昆虫体内是受调控的。这项工作将集中在控制上 在蚊子分子水平上的基因表达。一个很长的 这项研究的长期目标是为 允许对蚊子进行基因操作的技术。 在蚊子繁殖中发挥作用的基因提供了一个 发展分子生物学和遗传学的必要工具 这种重要的害虫。本提案的重点是 在卵子发育过程中表达的雌性特有基因 对类固醇激素20-羟基蜕皮激素的反应。我们计划 为了建立在我们已经确定的早期研究的基础上， 基因组文库，编码卵黄蛋白原基因的4个克隆-它们是 只在脂肪体内表达，对蜕皮激素的反应-和 另一个包含蜕皮激素诱导基因的克隆表达了 尤其是在卵泡细胞中。 这项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结构 赋予蜕皮激素反应性和基因的组织特异性 表情。由于对基因表达的功能分析 蚊子需要高效的转化技术， 它的发展受到严重阻碍，因为缺乏 克隆蚊子基因，这项提议的第二个主要目标是 优化蚊子基因转移系统，以实现快速分析 荷尔蒙诱导基因的体外构建。 很明显，蚊子的基因转化是一种 考虑到提供的强大工具，合理的目标 重组DNA技术，以及正在取得的快速进展 各种植物和动物的转变。vt.给出 蚊子明显的医学重要性，选择这种 昆虫作为基因转化的特定目标是 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.