Post Genomic Functional Genetic Analysis of Parasite/Insect Interaction

最近的生物学研究方向集中在对生物体的总DNA含量进行测序。这导致了潜在的所有基因编码的特定蛋白质存在于特定物种的识别。现在需要的是确定这些基因的功能，它们的蛋白质产物和产生它们的组织。建立基因功能最常见的方法是在试管中分离特定的基因，修改它们，然后将它们返回到生物体中。这些修饰可以导致生物体过量产生，或者相反，产生不足，这种特定的蛋白质，这反过来又导致特定的生理变化，提供有关基因正常作用的重要信息。在模式生物果蝇中，有许多系统可用于修饰基因表达。然而，要研究仅发生在寄生虫和吸血昆虫之间的重要相互作用，需要为这些重要的疾病携带者开发类似的方法。这与蚊子特别相关，蚊子是世界范围内的叮咬公害，也是许多人类疾病的最重要媒介。维持吸血昆虫的一般并发症，使有效的修改工具，最大限度地提高实验的灵活性的必要条件的发展。因此，我们将采用果蝇研究中最成功的方法来研究蚊子，冈比亚按蚊是非洲疟疾寄生虫的主要传播者。系统的不同组成部分将首先在分离的蚊子细胞的简化环境中进行测试，以快速识别那些功能最有效的蚊子细胞。然后将直接在蚊子中检查最有效的工具，以精确定义在整个生物体更复杂的整体环境中起作用的工具。为了证明该系统在蚊子中有效地发挥作用，我们将过量生产并沉默被认为可以杀死昆虫中寄生虫的特定蛋白质。利用该系统的灵活性，我们将确定如果蛋白质是由蚊子的“肝脏”、血细胞或胃产生的，疟疾寄生虫的杀灭是否更有效。虽然这项工作的重点是按蚊和疟原虫，但其原理和方法应可推广到由蚊子和吸血昆虫传播的其他寄生虫。

The recent direction of biological research has focused on sequencing the total DNA content of organisms. This has led to the identification of potentially all the genes encoding for specific proteins present in particular species. What is needed now is to identify the function of these genes, their protein products and the tissues in which they are produced. The most common way to establish gene function is to isolate specific genes in the test tube, modify them and then return them to the organism. These modifications can either cause the organism to over-produce, or conversely, underproduce, this specific protein which in turn causes a specific physiological change that gives vital information on the gene's normal role. In the model organism, the fruit fly, there are numerous systems available to modify gene expression. However, to study important interactions that only occur between parasites and blood sucking insects requires similar methods to be developed for these important carriers of disease. This is particular relevant to mosquitoes which are biting nuisances worldwide and the most important vectors of many human diseases. The general complications of maintenance of blood-feeding insects make the development of efficient modification tools that maximize experimental flexibility a requisite. We will thus adopt the most successful method used in fruit fly research to the mosquito, Anopheles gambiae that is the primary transmitter of the malaria parasite in Africa. Different components of system will be initially tested in the simplified environment of isolated mosquito cells to rapidly identify those that function most efficiently. A selection of the most efficient tools will then be examined directly in mosquitoes to precisely define those that work in the more complex holistic environment of the whole organism. As a proof of principle that the system functions efficiently in mosquitoes we will over-produce and silence specific proteins thought to kill parasites in the insect. Using the flexibility of the system we will determine whether malaria parasite killing is more effective if the protein is produced by the mosquitoes' 'liver', blood cells or its stomach. Although the work is focused on Anopheles mosquitoes and the malaria parasite, the principles and methods should be transferable to other parasites transmitted by mosquitoes and blood sucking insects.