THE COMPLETE GENOME SEQUENCE AND ANALYSIS OF M SMEGMATIS

两株M.实验室分离株H37Rv和临床分离株CDC 1551为研究结核分枝杆菌的感染性、毒力、定殖、致病性以及药物敏感性和耐药性机制提供了前所未有的资源。结核 尽管基因组序列提供了丰富的信息，但该生物极慢的生长速度和高毒性的性质使研究M。结核病及其基因操纵的艰巨任务。 在开发M.作为M.结核 通过这些努力，分枝杆菌研究人员有可能利用M。smegenerative在很大程度上与E.大肠杆菌用于更大的微生物群落。 获得M.耻垢病将是分枝杆菌研究的一个里程碑。 除了为继续发展其作为重组方法学的遗传系统提供宝贵的信息外，该生物体还为研究分枝杆菌生物学的独特方面提供了有价值的模型，包括细胞壁生物合成、生长速率调节、药物敏感性、蛋白质分泌、基因调节、基因组研究所已经开发了一种成本有效和高效的微生物基因组测序方法，该方法已经用于测序和组装七种微生物，基因组 分枝杆菌基因组将使用这种全基因组鸟枪法对耻垢病菌株MC 2155进行测序。 将制备小和大插入随机文库，并将获得足够数量的随机鸟枪序列以达到8倍覆盖。基因组的组装将使用TIGR组装软件的2.0版完成，并将使用该研究所开发的各种基于PCR的技术进行封闭。基因组的注释将采用多种计算机技术，用于识别开放阅读框架，并通过对策展数据库的相似性搜索来识别已知功能的蛋白质。 额外的分析将识别诸如输出蛋白质、tRNA和rRNA基因、重复序列、启动子序列和核糖体结合位点等特征。 这些数据将通过万维网（www.tigr.org）上的TIGR微生物数据库提供给研究界。

The completion and annotation of the DNA sequence from two strains of M. tuberculosis, the laboratory strain H37Rv and the virulent clinical isolate CDC1551 will provide the mycobacteria and tuberculosis research communities with an unprecedented resource for investigating the infectivity, virulence, colonization, pathogenicity, and the mechanisms of drug- sensitivity and resistance of M. tuberculosis. Despite the abundance of information provided by the genomic sequence, the exceedingly slow growth rate and highly virulent nature of the organism make working with M. tuberculosis and its genetic manipulation formidable tasks. A great deal of effort has gone into developing M. smegmatis, a fast-growing non-pathogen, as a genetic surrogate for M. tuberculosis. Through these efforts, it is possible for the mycobacterial researcher to utilize M. smegmatis in much the same manner as E. coli is used in the greater microbiological community. Obtaining the complete genome sequence of M. smegmatis will represent a milestone in mycobacterial research. In addition to providing invaluable information for continuing its development as a genetic system for recombinant methodologies, the organism provides a valuable model for studying unique aspects of mycobacterial biology, including cell wall biosynthesis, regulation of growth-rate, drug-sensitivity, protein secretion, gene regulation, etc. The Institute for Genomic Research has developed a cost effective and efficient approach to microbial genome sequencing that has been used to sequence and assemble seven microbial genomes. The genome of M. smegmatis strain mc2155 will be sequenced using this whole genome shotgun strategy. Small and large insert random libraries will be prepared and a sufficient number of random shotgun sequences to reach 8-fold coverage will be obtained. Assembly of the genome will be done using the version 2.0 of the TIGR Assembly software and closure will be obtained with a variety of PCR based techniques developed at The Institute. Annotation of the genome will employ a variety of computer techniques for identifying open reading frames and identifying proteins of known function through similarity searches against curated databases. Additional analyses will identify features such as exported proteins, tRNA and rRNA genes, repeated sequences, promoter sequences and ribosome binding sites. The data will be made available to the research community through the TIGR Microbial Database on the World Wide Web (www.tigr.org).