Analysis of Plasmodium development in the mosquito

描述：(申请人提供)：疟疾导致100多万人死亡 每年都有很多人。因为目前的药物干预程序， 杀虫剂和疫苗是无效的疟疾控制新方法 都是迫切需要的。疟疾的传播取决于 疟原虫寄生在蚊子的媒介中。然而，在 分子水平，对复杂的发育程序知之甚少 它控制着蚊子中疟原虫的分化。拟议中的项目 寻求缩小这种知识鸿沟。 在这个实验室进行的初步实验已经组装了许多 该项目所需的工具。已构建了四个cdna消减文库。 构建了富含在不同阶段表达的序列的 蚊子体内的疟原虫发育。包含4,000个克隆的微阵列 正在生产中，超过2,000个插入物正在测序。使用这些 工具，在特定阶段优先表达的疟原虫基因 将确定蚊子(而不是脊椎动物宿主)的发育。 并被选中进行进一步的研究。将特别关注基因 编码具有跨膜结构域的蛋白质(候选受体和 传播阻断抗原)或具有暗示参与 调节功能(例如，信号转导、激酶结构域、与 转录因子)。这些基因的时间表达将被研究 并将产生抗体来确定细胞的定位。这个 这些抗体可能阻止寄生虫在体内的发展 将对蚊子进行评估。基因敲除突变会产生和改变 由这些突变引起的基因表达将通过使用 微阵列。

DESCRIPTION: (provided by the applicant): Malaria kills over one million persons every year. Because current intervention procedures with drugs, insecticides and vaccines are ineffective, new approaches for malaria control are urgently needed. Malaria transmission depends on the ability of the Plasmodium parasite to develop in the mosquito vector. However, at the molecular level, very little is known about the complex developmental program that governs Plasmodium differentiation in the mosquito. The proposed project seeks to reduce this knowledge gap. Initial experiments conducted in this laboratory have assembled many of the required tools for the project. Four cDNA subtraction libraries have been constructed that are enriched for sequences expressed at different stages of Plasmodium development in the mosquito. Micro-arrays containing 4,000 clones are being produced and over 2,000 inserts are being sequenced. Using these tools, Plasmodium genes that are preferentially expressed at specific stages of development in the mosquito (and not in the vertebrate host) will be identified and selected for further study. Particular attention will be devoted to genes encoding proteins with transmembrane domains (candidate receptors and transmission-blocking antigens) or that have motifs suggesting involvement in regulatory functions (e.g., signal transduction, kinase domains, similarity to transcription factors). Temporal expression of these genes will be investigated and antibodies will be produced to determine cellular localization. The possible ability of these antibodies to block development of the parasite in the mosquito will be assessed. Knockout mutations will be produced and changes of gene expression caused by these mutations will be measured by use of the micro-arrays.