GENES FOR T CRUZI KINETOPLAST DNA ASSOCIATED PROTEINS

动质体目的成员，包括锥虫和 利什曼原虫是主要的热带疾病。 这些 有鞭毛的原生动物拥有最不寻常的线粒体DNA之一 在自然界中发现的结构。 这些细胞中的单个线粒体 生物体含有相对大比例的细胞DNA， 动质体DNA（kDNA），以盘状结构组织， 两类拓扑相连的圆形种群的上界 分子。 尽管最近我们在理解上取得了进展， kDNA复制，关于这个复杂的DNA的悬而未决的问题仍然存在 结构我们研究的主要目标是为更好地 了解kDNA网络的结构和功能。我们希望 认为特定的蛋白质-kDNA相互作用在 这些结构的维护和隔离。因此 kDNA相关蛋白（KAPS）的鉴定和表征 可能会提供一些关于这些不寻常的DNA功能的信息， 结构. 我们把注意力集中在一组非组蛋白样的 在2 M后仍与克氏锥虫kDNA结合的蛋白质 在温和洗涤剂存在下NaCl提取kDNA。 使用 免疫电镜和荧光显微镜观察到 兔子的抗血清再次激活了这些蛋白质， 定位于T. cruzi细胞 更有趣的是， 相同的抗体制剂与相似定位的表位交叉反应 亚马逊利什曼原虫和布氏锥虫。 基于这些 根据观察，本研究建议试图确定和 描述T.克鲁兹KAPS。 我们建议首先筛选一个T。cruzi cDNA 表达文库与上述抗体制备物， 第二，筛选和鉴定编码T. cruzi KAPs。通过对T. cruzi KAPs我们 可以学到很多关于kDNA网络是如何维持的， 传播。 这些信息也可能为我们提供一个更合理的 彻底根除T. cruzi病原体 美洲锥虫

Members of the order Kinetoplastida which include Trypanosoma and Leishmania are responsible for major tropical diseases. These flagellated protozoa share one of the most unusual mitochondrial DNA structures found in nature. The single mitochondria present in these organisms contains a relatively large proportion of the cellular DNA, kinetoplast DNA (kDNA), organized in a disk-shaped structure and made up of a population of two types of topologically linked circular molecules. In spite of recent advances made in our understanding of kDNA replication, outstanding questions remain about this complex DNA structure. The major goal of our research is to contribute to a better understanding about the structure and function of kDNA networks. We wish to argue that specific protein-kDNA interactions play a key role in the maintenance and segregation of these structures. Therefore, the identification and characterization of kDNA-associated proteins (KAPS) may provide some information about the function of these unusual DNA structures. We have focused our attention on a set of non-histone-like proteins that remain associated to Trypanosoma cruzi kDNA after a 2M NaCl extraction of kDNA in the presence of a mild detergent. Using immuno-electron microscopy and fluorescent microscopy we have observed that rabbit antisera raised again these proteins, recognize epitopes localized in the kinetoplast of T. cruzi cells. More interestingly, the same antibody preparation cross-reacts with similarly localized epitopes in Leishmania amazonensis and Trypanosoma brucei. Based on these observations, the present research proposal attempts to identify and characterize T. cruzi KAPS. We propose first, to screen a T. cruzi cDNA expression library with the antibody preparation described above and second, to select and characterize clones encoding putative T. cruzi KAPs. By understanding the structure and function of T. cruzi KAPs we may learn a great deal about how kDNA networks are maintained and propagated. This information may also provide us with a more rational approach toward the complete eradication of T. cruzi the causative agent of American trypanosomiasis.