COMPLETE SEQUENCING AND GENE EXPRESSION IN M AVIUM

M AVIUM 中的完整测序和基因表达

• 批准号: 2887594
• 负责人: ROBERT D FLEISCHMANN
• 金额: $ 63.29万
• 依托单位: INSTITUTE FOR GENOMIC RESEARCH
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2887594
• 关键词: Mycobacterium avium; bacterial genetics; gene expression; genetic library; microorganism culture; microorganism growth; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction

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.

描述：（改编自申请人的摘要）该项目的目标 确定鸟分枝杆菌的完整 DNA 序列 染色体并用各种方法检查基因表达的全局变化 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得到证实 最近由 P.I.和他的研究所。 将使用的方法是 以前用于流感嗜血杆菌等的那些，尽管基因组较大 大小（流感嗜血杆菌为 4.7 Mbp，而流感嗜血杆菌为 1.8）则需要近 3 倍 扩大规模。 将进行序列的标准分析。 检查 基因表达的变异，估计 4500 个基因的 PCR 产物将 排列在滤膜上，与从 RNA 制备的标记 cDNA 杂交 在估计 27 种不同条件下生长的 M. avium （涉及生长阶段、养分和氧气消耗、温度、pH、 自由基、抗菌剂和巨噬细胞内的生长）。 这 申请人相信完整的 M. avium 序列，连同 目前正在确定完整的结核分枝杆菌和麻风分枝杆菌序列， 应该可以通过以下方式对基因组和基因进行信息丰富的比较 基因基础。 该序列将在疫苗方面具有潜在的应用 开发、毒力机制鉴定和抗菌药物 治疗和抵抗力。