MOLECULAR ANALYSIS OF THE MOSQUITO PERITROPHIC MATRIX

蚊子周食基质的分子分析

• 批准号: 6782312
• 负责人: MARCELO JACOBS-LORENA
• 金额: $ 27.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6782312
• 关键词: Anopheles; Plasmodium berghei; antibody; chimeric proteins; chitin; complementary DNA; disease vectors; extracellular matrix proteins; gastrointestinal epithelium; gene expression; genetic library; genetic regulatory element; host organism interaction; laboratory mouse; malaria; molecular cloning; northern blottings; nucleic acid sequence; polymerase chain reaction; protein sequence; protein structure function; structural genes; western blottings

Mosquito-transmitted diseases are a major cause of suffering and death in the tropical world. Malaria alone, kills one to two million people every year. The urgency for developing new control strategies is underscored by the development of resistance by parasites to previously effective drugs, by the resistance of mosquitoes to a variety of insecticides, and by the lack of an effective vaccine. Inhibition of the parasite's life cycle in the mosquito is a strategy that requires more attention. This proposal focuses on the gut of the human malaria vector, Anopheles gambiae. The gut is the first site of interaction between Plasmodium and the mosquito. Ingestion of blood by the adult mosquito triggers the secretion of a peritrophic matrix (PM), which is a thick extra-cellular sheath that completely surrounds the blood meal and any ingested parasites. The PM poses a partial barrier for malaria parasite invasion. Modifications of the PM may lead to a more complete barrier to infection. To devise such strategies will require a thorough molecular characterization of the PM, and a more complete understanding of its structure and function. The major objectives of this proposal are to isolate and characterize genes encoding PM proteins, to investigate how PM proteins interact for the assembly of the PM, and to probe into the physiological role of the PM. Specifically, PM genes will be cloned by screening expression libraries with anti-PM antibodies or based on amino acid sequence of fractionated PM proteins. Interaction between PM proteins will be measured in vivo using the yeast two-hybrid system or in vitro using affinity blotting techniques. Antibodies to recombinant PM proteins will be used to determine if PM proteins are stored in secretion vesicles. The thickness and porosity of the PM will be experimentally altered to measure the effects on digestion and on the ability of parasites to traverse the PM. Genetic transformation of Ae. aegypti is already possible and transformation of An. gambiae is likely to become available in the near future. The characterization of PM protein genes and an understanding of their function may lead to novel strategies for malaria control.

蚊子传播的疾病是热带世界中遭受痛苦和死亡的主要原因。仅疟疾，每年就会杀死1-200万人。寄生虫对以前有效的药物，蚊子对多种杀虫剂的耐药性以及缺乏有效的疫苗的抗性，迫切需要开发新的控制策略的紧迫性。抑制寄生虫在蚊子中的生命周期是一种需要更多关注的策略。该提议着重于人类疟疾载体Anopheles Gambiae的肠道。肠道是疟原虫和蚊子之间相互作用的第一个部位。成年蚊子摄入血液会引发营养基质（PM）的分泌，该基质是一种厚细胞外护套，完全围绕着血液粉和任何摄入的寄生虫。 PM对疟疾寄生虫入侵构成了部分障碍。 PM的修改可能会导致更完整的感染障碍。为了制定这种策略，将需要对PM进行彻底的分子表征，并对其结构和功能有更完整的了解。该提案的主要目标是分离和表征编码PM蛋白的基因，以研究PM蛋白如何与PM组装相互作用，并探测PM的生理作用。具体而言，PM基因将通过筛选抗PM抗体或基于分级PM蛋白的氨基酸序列来克隆。 PM蛋白之间的相互作用将使用酵母两杂交系统或使用亲和力印迹技术在体内测量。重组PM蛋白的抗体将用于确定PM蛋白是否存储在分泌囊泡中。 PM的厚度和孔隙率将进行实验改变，以测量对消化的影响以及寄生虫穿越PM的能力。 AE的遗传转化。埃及已经是可能的，也是An的转变。冈比亚很可能会在不久的将来上市。 PM蛋白基因的表征及其对其功能的理解可能会导致疟疾控制的新策略。