COMPLETE SEQUENCING AND GENE EXPRESSION IN M AVIUM

描述：（改编自申请人摘要）本项目的目标 确定鸟分枝杆菌的完整DNA序列 染色体和检查基因表达的全球变化与各种 体外环境条件的变化。 可行性和 微生物基因组测序的成本效益已经得到证明 最近被私家侦探他的研究所。 将采用的方法是 以前用H。流感等，尽管较大的基因组 大小（4.7Mbp对H.流感）将需要近3倍的 按比例放大。 将对序列进行标准分析。 审查 基因表达的变异，估计4500个基因的PCR产物将 排列在过滤器上，用于与从RNA制备的标记cDNA杂交 分枝在大约27种不同的条件下生长的鸟类 （涉及生长阶段、营养物和氧气消耗、温度、pH、 自由基、抗微生物剂和巨噬细胞内的生长）。 的 申请人认为，完整的M. avium序列，结合 完全M.结核和M.麻风病序列目前正在确定， 应该导致基因组和基因的高度信息化比较， 基因基础 该序列在疫苗方面具有潜在的应用价值 毒力机制的开发、鉴定和抗菌 治疗和抵抗

DESCRIPTION: (Adapted from Applicant,s Abstract) The goals of this project are to determine the complete DNA sequence of the Mycobacterium avium chromosome and examine global changes in gene expression with various changes in environmental conditions in vitro. The feasability and cost-effectiveness of microbial genome sequencing has been demonstrated recently by the P.I. and his institute. The approaches to be used will be those used previously with H. influenzae, etc., although the larger genome size (4.7 Mbp versus 1.8 for H. influenzae) would require a nearly 3 fold scaleup. Standard analyses of the sequence will be conducted. To examine variation in gene expression, PCR products of the estimated 4500 genes will be arrayed on filters for hybridization to labelled cDNA prepared from RNA of M. avium grown under an estimated 27 different sets of conditions (involving growth phase, nutrient and oxygen depletion, temperature, pH, radicals, antimicrobial agents, and growth within macrophages). The applicant believes that complete M. avium sequence, in conjuction with the complete M. tuberculosis and M. leprae sequences currently being determined, should lead to highly informative comparisons on both a genome and gene by gene basis. The sequence will have potential applications to vaccine development, identification of virulence mechanisms, and antimicrobial therapy and resistance.