Identification of E. histolytica virulence factors

描述(申请人提供)：腹泻病每十人中就有一人死于腹泻 在孟加拉国等贫穷国家，儿童在他或她五岁生日之前出生。 溶组织内阿米巴是该病的病原菌之一。这是一种人类寄生虫 具有非凡的入侵和裂解宿主组织的能力。侵入性 感染溶组织埃希氏菌会导致痢疾、阿米巴结肠炎和阿米巴。 肝脓肿。内阿米巴基因组计划即将完成 提供了一个前所未有的机会将遗传学的方法应用于 阿米巴病的问题。提出了三个具体目标：(1)表征 宿主：通过识别招募到这个位置的分子来识别寄生虫的界面 本发明公开了一种利用酵母双杂交系统，(2)使用一种 蛋白质组学方法与溶组乳杆菌基因组测序的结合 数据库进一步鉴定与GalNAc相互作用的新蛋白质 凝集素，以及(3)研究溶组织乳杆菌的生物学意义 被GalNAc凝集素招募到宿主的蛋白质：寄生虫通过 使用干扰RNA(RNAi)创建这些蛋白质的“敲除”突变体， 反义RNA技术和/或显性负性突变体的表达。 成功完成这些研究将产生一个根本性的理解 与溶组乳杆菌GalNAc凝集素相互作用的阿米巴蛋白 它们在致病性中的作用。

DESCRIPTION (provided by applicant): Diarrheal diseases kill one of every ten children by his or her fifth birthday in poor countries such as Bangladesh. Entamoeba histolytica is one of the etiologic agents. It is a human parasite that has a remarkable ability to invade and lyse host tissues. Invasive infection with E. histolytica leads to dysentery, amebic colitis, and amebic liver abscess. The imminent completion of the Entamoeba genome project provides an unprecendented opportunity to apply the approaches of genetics to the problem of amebiasis. Three specific aims are proposed: (1) Characterize the host:parasite interface by identifying molecules recruited to this site by the E. histolytica GalNAc lectin using a yeast two-hybrid system, (2) Use a proteomics approach in conjunction with the E. histolytica genome sequencing database to further characterize novel proteins that interact with the GalNAc lectin, and (3) Investigate the biological significance of E. histolytica proteins recruited by the GalNAc lectin to the host:parasite interface by creating "knockout" mutants of these proteins using interference RNA (RNAi), antisense RNA technology, and/or expression of dominant negative mutants. Successful completion of these studies will yield a fundamental understanding of amebic proteins that interact with the E. histolytica GalNAc lectin and their role in pathogenicity.