AFRICAN TRYPANOSOME GENOME SEQUENCING

非洲锥虫是一种原生动物寄生虫， 人类的昏睡病和家畜的恩加纳病。 在过去的二十年里，估计有2.38亿美元被用于 在全世界范围内用于非洲锥虫的基础研究。 这 研究不仅使我们对这些疾病的理解有了很大的进步， 这些生物，还有一些以前未知的生物， 现象，其中一些后来被发现发生在细胞中 包括哺乳动物在内的其他生物。 然而，这些重要的研究 但这些努力并没有带来更好的方式来管理或控制这一点 毁灭性的疾病最近，人类基因组计划的进展， 酵母基因组序列的测定， 证明了使用基因组序列作为基础的价值， 设计未来的研究工作。 我们的长期目标是 布氏锥虫的基因组 这项建议是实现这一目标的第一步，我们 提出以下战略，分两个阶段。 上 阶段，我们将产生约20 Mb的不连续单程 序列（27 Mb非微型染色体基因组的74%）。 这 将通过对10，000个已经存在的克隆进行末端测序来实现 （目前正在用于组装T.布鲁氏菌分离株 TREU 927/4DNA），5000个BAC克隆（目前由Sara梅尔维尔， 剑桥大学，UK）和5000个随机- 剪切的DNA（由TIGR构建）。端粒近端序列 含有端粒连接的VSGs将在 剪切的基因组文库。 第一阶段的目的是加强 早期的基因发现，并提供标记，将是重要的， 构建高分辨率序列就绪图。 在第二 相，10 Mb的连续T.布氏杆菌DNA将使用 一个接一个染色体的方法该项目将提供宝贵的 在许多层面上的信息和好处，包括（1）识别 真核细胞的基本功能中涉及的基因，（2）容易， 廉价和快速克隆编码蛋白质的基因正在积极地 在世界各地的实验室进行研究，（3）立即获得基因 及其功能/结构研究的产物，（4）预测 代谢途径的候选基因的基础上，和（5） 通过比较鉴定寄生虫特异性基因项目 其他基因组

African trypanosomes are protozoan parasites that cause a fatal disease called sleeping sickness in humans and ngana in domestic livestock. During the past two decades, an estimated 238 million dollars has been spent worldwide on basic research on African trypanosomes. This research has resulted in many advances in our understanding of not only these organisms, but also a number of previously unknown biological phenomena, some of which have been subsequently found to occur in cells of other organisms including mammals. Yet, these substantial research efforts have not led to a better way to manage or control this devastating disease. Recently, progress on the human genome project, and the determination of the yeast genomic sequence, have unambiguously demonstrated the value of using genomic sequences as a foundation for designing future research efforts. Our long term goals is to sequence the genome of Trypanosoma brucei. This proposal represents the first step towards that goal, and we propose the following strategy consisting of two phases. In the first phase, we will generate about 20 Mb of discontinuous single-pass sequence (74 percent of the 27 Mb non-minichromosomal genome). This will be implemented by end-sequencing of 10,000 already-existing clones (currently being used to assemble the physical map of T. brucei isolate TREU 927/4DNA), 5000 BAC clones (currently being made by Sara Melville, Cambridge Univ., UK) and 5000 small insert plasmid clones of randomly- sheared DNA (to be constructed by TIGR). Telomere-proximal sequences containing the telomere-linked vsgs will be identified in clones of the sheared genomic library. The purpose of this first phase is to enhance early gene discovery and to provide markers that will be important for construction of a high-resolution sequence-ready map. During the second phase, 10 Mb of contiguous T. brucei DNA will be sequenced using a chromosome by chromosome approach. This project will provide invaluable information and benefits at many levels, including (1) identification of genes involved in basic functions of the eukaryotic cells, (2) easy, inexpensive and fast cloning of genes encoding proteins being actively studied in laboratories around the world, (3) immediate access to genes and their products from functional/structural studies, (4) prediction of metabolic-pathways on the basis of candidate genes, and (5) identification of parasite-specific gene projects by comparison with other genomes.