Chuang S E, Daniels D L, Blattner F R
Laboratory of Genetics, University of Wisconsin-Madison 53706.
J Bacteriol. 1993 Apr;175(7):2026-36. doi: 10.1128/jb.175.7.2026-2036.1993.
Global transcription responses of Escherichia coli to various stimuli or genetic defects were studied by measuring mRNA levels in about 400 segments of the genome. Measuring mRNA levels was done by analyzing hybridization to DNA dot blots made with overlapping lambda clones spanning the genome of E. coli K-12. Conditions examined included isopropyl-beta-D-thiogalactopyranoside (IPTG) induction, heat shock, osmotic shock, starvation for various nutrients, entrance of cells into the stationary phase of growth, anaerobic growth in a tube, growth in the gnotobiotic mouse gut, and effects of pleiotropic mutations rpoH, himA, topA, and crp. Most mapped genes known to be regulated by a particular situation were successfully detected. In addition, many chromosomal regions containing no previously known regulated genes were discovered that responded to various stimuli. This new method for studying globally regulated genetic systems in E. coli combines detection, cloning, and physical mapping of a battery of coregulated genes in one step.
通过测量大肠杆菌基因组约400个区段中的mRNA水平,研究了大肠杆菌对各种刺激或基因缺陷的全局转录反应。通过分析与用跨越大肠杆菌K-12基因组的重叠λ克隆制成的DNA斑点印迹的杂交来测量mRNA水平。所研究的条件包括异丙基-β-D-硫代半乳糖苷(IPTG)诱导、热休克、渗透休克、各种营养物质饥饿、细胞进入生长稳定期、在试管中厌氧生长、在无菌小鼠肠道中生长以及多效性突变rpoH、himA、topA和crp的影响。大多数已知受特定情况调控的定位基因都被成功检测到。此外,还发现了许多不含先前已知调控基因的染色体区域对各种刺激有反应。这种研究大肠杆菌全局调控遗传系统的新方法将一组共调控基因的检测、克隆和物理定位一步完成。