T CRUZI GENOME: UU

克氏锥虫是一种引起恰加斯病的原生动物寄生虫，影响数百万人，构成南美洲和中美洲的主要健康问题。 目前还没有有效的药物来对抗这种疾病，也没有开发出疫苗，尽管已经对T.克鲁兹几十年来 在过去的几年里，在各种基因组计划中产生的大量有用的数据已经证明了这种方法在基因发现和基因组表征方面的能力。 因此，在过去三年中，开展了基因组绘图和测序试点项目，以提高基因发现的效率，从而确定潜在的药物靶点，并为今后的基因组工作建立知识基础。 这些项目包括初步核型分析、几条染色体的绘图、EST测序和3号染色体的测序。 后一个项目以及绘图和EST测序已在该实验室进行。 为了利用T. cruzi已经积累，我们在这里建议，与TIGR（Najib El-Sayed）和SBRI（Ken Stuart）的团队一起，发起一项大规模的工作来描述T。cruzi基因组 选择的菌株将是初始项目中使用的CL Brener参考菌株。 该生物体的单倍体基因组大小约为43.5 Mb。本项目的目标是：（1.）通过将1，000个特异性标记与分离的染色体杂交来表征该菌株的核型，这将产生每40-50 kb具有一个标记的连锁图。 （2.）从基因组BAC克隆进行末端测序，以建立物理图谱和用于基因发现。 （3.）T. cruzi CL Brener使用基于从（2.）大插入克隆末端测序和核型分析将在提案的第一年进行，第2年和第3年将致力于逐个染色体测序。 我们计划在第2年和第3年开展核型分析项目，并参与测序工作，对7，000个BAC克隆进行末端测序，并完成10-15 Mb基因组DNA的测序。 该项目将产生大量关于药物和疫苗潜在目标的数据，并将与其他寄生虫基因组项目的数据一起成为全世界寄生虫学家的极其宝贵的资源。 我们预见，这个项目将带来大量的调查基因功能，基因表达，进化和功能研究的基础上计算生物学方法，并测试潜在的药物靶点。

Trypanosoma cruzi is a protozoan parasite that causes Chagas' disease, which affects several million people and constitutes a major health problem in South and Central America. No effective drug is available against this disease and no vaccine has been developed, even though basic research has been carried out into the biology of T. cruzi for decades. The large amount of useful data produced in the last years within various genome projects have demonstrated the power of this approach for gene finding and genome characterization. Therefore, in the last three years, pilot genome mapping and sequencing projects have been carried out to increase the efficiency of gene finding to identify potential drug targets and to create a knowledge base for future genome efforts. These projects have included initial karyotyping, mapping of several chromosomes, EST sequencing and sequencing of chromosome 3. The latter project as well as mapping and EST sequencing has been carried out in this laboratory. In order to take advantage of the knowledge of the genome organization of T. cruzi that has been accumulated, we here propose, together with groups at TIGR (Najib El-Sayed) and SBRI (Ken Stuart), to initiate a large effort to characterize the T. cruzi genome. The strain of choice will be the CL Brener reference strain that has been used in the initial projects. The haploid genome size of this organism is approximately 43.5 Mb. The goals of the project are: (1.) to characterize the karyotype of this strain by hybridization of 1,000 specific markers to separated chromosomes, which will result in a linkage map with a marker every 40-50 kb. (2.) to carry out end-sequencing from genomic BAC- clones in order to establish a physical map and for gene discovery. (3.) sequencing of chromosomes in T. cruzi CL Brener using clones selected on the basis of the marker and end-sequence information obtained from all three laboratories in (2.) Large-insert clone end sequencing and karyotyping will be carried out in the first year of the proposal and years 2 and 3 will be dedicated to chromosome-by-chromosome sequencing. We propose to carry out the karyotyping project as well as participate in the sequencing effort by carrying out end-sequencing from 7,000 BAC clone and complete sequencing of 10-15 Mb of genomic DNA in years 2 and 3. This project will result in a wealth of data on potential targets of drugs and vaccines and will, together with data from other parasite genome projects, be an extremely valuable resource for parasitologists world wide. We foresee that this project will bring about a plethora of investigations into gene function, gene expression, evolutionary and functional studies based on computational biology methods, and testing of potential drug targets.