Molecular repertoire of rhoptry organelles from Plasmodium falciparum

镰状体是恶性疟原虫子体中配对的分泌细胞器，被认为在疟原虫红细胞阶段的宿主细胞入侵中起关键作用。关于顶复合体的细胞器的功能，特别是关于顶复合体的细胞器，以及它们的关键成分的知识是很少的。拟议项目的主要问题是对蛇体的分子库进行分析，即鉴定和表征蛇体蛋白质，其中大部分仍是未知的。我们将通过差速离心富集植物细胞器，并通过二维凝胶电泳检查它们的蛋白质含量，这是一种已经变得复杂的技术，对生物体蛋白质组的分析特别有吸引力。“状体蛋白组”分离后，将对几个突出的蛋白点进行n端测序。新鉴定的蛋白质将被分子克隆以阐明其初级结构，并将序列与数据库中的其他蛋白质进行比较。重组虫体蛋白的表达和产生针对这些因子的特异性抗体将有助于功能表征它们，并阐明虫体细胞器及其关键成分在寄生虫入侵中的作用。

Rhoptries are paired secretory organelles in Plasmodium falciparum merozoites and are suggested to play a crucial role in host cell invasion during erythrocytic stages of the malaria parasite. The knowledge about the function of the organelles of the apical complex, particularly of rhoptries, and about their key components is scarce. The major issue of the proposed project is the analysis of the molecular repertoire of rhoptries, i.e. the identification and characterization of rhoptry proteins, the majority of which is still unknown. We will enrich rhoptry organelles by differential centrifugationand examine their protein content by Two-dimensional gel electrophoresis, a technique which has become sophisticated andparticularly attractive for the analysis of proteoms of organisms. After separation of the "rhoptry proteom", severalprominent protein spots will be subjected to N-terminal sequencing. Newly identified proteins will be molecularly cloned to elucidate their primary structures, and the sequenceswill be compared with those of other proteins in data bases. Recombinant expression of selected rhoptry proteins and generation of specific antibodies against these factors will help to functionally characterize them and to elucidate the role of rhoptry organelles and their key components in parasiteinvasion.