T CRUZI GENOME: UU

克氏锥虫是一种引起查加斯病的原生动物寄生虫，影响着数百万人，构成了中南美洲的一个主要健康问题。尽管对克氏锥虫的生物学进行了数十年的基础研究，但目前还没有针对这种疾病的有效药物，也没有开发出疫苗。过去几年在各种基因组计划中产生的大量有用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力量。因此，在过去三年中，开展了试验性基因组测绘和测序项目，以提高基因发现的效率，以确定潜在的药物靶点，并为未来的基因组工作建立知识库。这些项目包括初步核型分析、几条染色体的定位、EST测序和3号染色体的测序。第三个项目以及作图和EST测序已经在本实验室进行。为了利用已经积累的关于克鲁兹毛滴虫基因组组织的知识，我们建议与TIGR(Najib El-Sayed)和SBRI(Ken Stuart)的团队一起发起一项大规模的努力来表征克鲁兹毛滴虫的基因组。选择的菌株将是已在初始项目中使用的CL Brener参考菌株。该生物的单倍体基因组大小约为43.5Mb。该项目的目标是：(1)通过将1,000个特定标记与分离的染色体杂交来鉴定该菌株的核型，这将导致每40-50 kb一个标记的连锁图谱。(2)对基因组BAC克隆进行末端测序，以建立物理图谱并用于基因发现。(3)根据(2)中从所有三个实验室获得的标记和末端序列信息，使用所选择的克隆对克氏锥虫的染色体进行了测序。大插入克隆末端测序和核型分析将在提议的第一年进行，第二年和第三年将专门用于逐条染色体的测序。我们计划在第二年和第三年对7000个BAC克隆进行末端测序，并完成10-15Mb的基因组DNA测序，从而实施核型测定计划并参与测序工作。这一计划将产生大量关于药物和疫苗潜在靶标的数据，并将与其他寄生虫基因组计划的数据一起，成为世界各地寄生虫学家极其宝贵的资源。我们预计，该项目将带来大量关于基因功能、基因表达、基于计算生物学方法的进化和功能研究以及潜在药物靶点的测试。

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.