GENOME ANALYSIS OF THE GENUS BDELLOVIBRIO

蛭弧菌属的基因组分析

• 批准号: 2208948
• 负责人: JOHN J TUDOR
• 金额: $ 11.15万
• 依托单位: SAINT JOSEPH'S UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-01 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2208948
• 关键词: DNA footprinting; Vibrio; bacterial genetics; gene expression; genetic manipulation; genetic mapping; genetic strain; genome; molecular cloning; pulsed field gel electrophoresis; restriction fragment length polymorphism; southern blotting; transposon /insertion element

Little is known about the genetics of the bdellovibrios, and a system for introducing DNA into bdellovibrios has only recently been reported. The development of a map of the bdellovibrio chromosome would serve as a stimulus for additional genetic studies and a framework onto which genetic data could be coordinated. The overall objectives are to construct a physical map of the Bdellovibrio bacteriovorus genome, to compare the physical maps of several bdellovibrio strains, and to locate a number of genes on the map. High quality, unsheared genomic DNA will be extracted from Bdellovibrio bacteriovorus 109J, and several other strains, into agarose beads. The DNA will be cut with rare cutting restriction endonucleases, and the resulting large DNA fragments separated using pulsed field gel electrophoresis. The restriction patterns, or fingerprints, of various strains will be compared in order to determine relatedness and taxonomic significance. Restriction fragments will be ordered relative to each other. This will be accomplished by 1)using fragments of one enzyme digest to probe Southern blots of different enzyme restriction patterns to discover overlaps between fragments; 2)probing Bdellovibrio genomic YAC clones with isolated genomic restriction fragments. The beginnings of a genetic map will be originated by positioning cloned genes and transposon insertions on the physical map.

人们对蛭弧菌的遗传学知之甚少， 将DNA引入蛭弧菌只是最近才有报道。 的 蛭弧菌染色体图谱的开发将作为 刺激更多的遗传研究和框架， 基因数据可以进行协调。 总体目标是 构建噬菌蛭弧菌基因组的物理图谱， 比较几种蛭弧菌菌株的物理图谱， 在地图上的一些基因。 高质量的未剪切的基因组DNA将 可以从噬菌蛭弧菌109J和其他几种噬菌蛭弧菌中提取 菌株，放入琼脂糖珠中。 DNA会被罕见的切割 限制性内切酶，以及产生的大DNA片段 使用脉冲场凝胶电泳分离。 的限制 各种菌株的模式或指纹将按顺序进行比较 以确定亲缘关系和分类学意义。 限制 片段将相对于彼此排序。 这将是 通过1）使用一种酶消化物的片段来探测Southern杂交来完成 不同酶限制性图谱的印迹以发现重叠 2）用探针探测蛭弧菌基因组YAC克隆， 分离的基因组限制性片段。 基因图谱的起源 将通过定位克隆基因和转座子插入 在物理地图上。