MOLECULAR GENETICS OF LEISHMANIA

锥虫原虫属利什曼原虫的寄生虫是 对一系列影响超过10个国家的热带疾病负责 全世界有数百万人，取决于特定的物种和免疫力 感染者的状态，可以是严重的或致命的。在巴西、非洲和 在地中海盆地，利什曼病是一种常见的机会性感染 艾滋病患者。这项提案的目标是开发基因治疗方法。 基因和蛋白质的识别可能在这一过程中发挥重要作用 原生动物寄生虫利什曼原虫的感染循环。前提是 强大的分子遗传学工具的可用性，以及基因的鉴定 对寄生虫的毒力很重要，将从根本上提高我们发展的能力 改进了控制策略。在以前的研究中，方法被开发用于 以各种方式操纵寄生虫基因组，并执行 功能性基因拯救。这导致了几位候选人的确定 毒力基因。UBF]等基因在寄生虫DNA中的作用 修复对生存至关重要的道路，将继续进行。随着……的快速发展 利什曼原虫基因组计划，以及最近在开发利什曼原虫基因组计划方面的突破 适用于全基因组寄生虫基因表达调查的新方法 启动一系列“功能基因组”研究的时机是合适的 以描述完整寄生虫基因组的表达为导向的。 我们的目标是找出显示出表达模式的基因 在感染周期中的功能作用，例如阶段特异性表达 或对相关刺激作出反应的改变。我们将使用基于水手的 用于系统地产生GFP基因融合的转位系统 体外的寄生虫基因组片段或体内的整个基因组。这些 方法的优点是在蛋白质水平上研究表达， 允许核酸和蛋白质的调节的累积效应 要评估的级别。有希望的DNA微阵列初步数据已经 并对利什曼原虫基因表达进行了一系列调查 在各种相关的发育阶段和环境处理下 将会被执行。

Parasites of the trypanosomatid protozoan genus Leishmania are responsible for a spectrum of tropical diseases that affect more than 10 million people worldwide, and depending on the specific species and immune status of the infected person, can be severe or fatal. In Brasil, Africa and the Mediterranean basin, leishmaniasis is a common opportunistic infection in AIDS patients. The goal of this proposal is to develop methods for the genetic identification of genes and proteins likely to play important roles in the infectious cycle of the protozoan parasite Leishmania. The premise is that the availability of powerful molecular genetic tools, and identification of genes important to parasite virulence, will radically advance our ability to develop improved control strategies. In previous studies methods were developed for manipulating the parasite genome in a variety of ways, and performing functional genetic rescue. These led to the identification of several candidate virulence genes. The role of genes such as UBF], implicated in parasite DNA repair pathways essential for survival, will be pursued. With rapid progress in the Leishmania genome project, and recent breakthroughs in the development of new methods suitable for genome-wide surveys of parasite gene expression, the time is appropriate to initiate a series of 'functional genomic' studies oriented towards characterizing expression from the complete parasite genome. The goal is to identify genes showing patterns of expression suggestive of functional roles in the infectious cycle, for example stage-specific expression or alterations in response to relevant stimuli. We will use mariner-based transpositional systems to systematically generate GFP gene fusions in defined segments of the parasite genome in vitro or the entire genome in vivo. These methods have the advantage of studying expression at the protein level, allowing the cumulative effect of regulation at both nucleic acid and protein levels to be evaluated. Promising preliminary data with DNA microarrays have been gathered, and a series of investigations of Leishmania gene expression under a variety of relevant developmental stages and environmental treatments will be carried out.