通过随机扩增多态性DNA分析检测大肠杆菌H10407中渗透胁迫诱导的基因重排。
Osmotic stress-induced genetic rearrangements in Escherichia coli H10407 detected by randomly amplified polymorphic DNA analysis.
作者信息
Jolivet-Gougeon A, David-Jobert S, Tamanai-Shacoori Z, Ménard C, Cormier M
机构信息
Laboratoire de Microbiologie Pharmaceutique, Université de Rennes I, 35000 Rennes, France.
出版信息
Appl Environ Microbiol. 2000 Dec;66(12):5484-7. doi: 10.1128/AEM.66.12.5484-5487.2000.
Abstract
Randomly amplified polymorphic DNA (RAPD) analysis is a DNA polymorphism assay commonly used for fingerprinting genomes. After optimizing the reaction conditions, samples of Escherichia coli H10407 DNA were assayed to determine the influence of osmotic and/or oligotrophic stress on variations in RAPD banding patterns. Genetic rearrangements or DNA topology variations could be detected as changes in agarose gel electrophoresis banding profiles. A new amplicon generated using DNA extracted from bacteria prestarved by an osmotic stress and resuscitated in rich medium was observed. Enrichment improved recovery of mutator cells and allowed them to be detected in samples, suggesting that DNA modifications, such as stress-induced alterations and supercoiling phenomena, should be taken into consideration before beginning RAPD analyses.
摘要
随机扩增多态性DNA（RAPD）分析是一种常用于基因组指纹分析的DNA多态性检测方法。在优化反应条件后，对大肠杆菌H10407 DNA样本进行检测，以确定渗透和/或贫营养胁迫对RAPD条带模式变化的影响。遗传重排或DNA拓扑结构变化可通过琼脂糖凝胶电泳条带图谱的变化来检测。观察到使用从经渗透胁迫预饥饿并在丰富培养基中复苏的细菌中提取的DNA产生了一个新的扩增子。富集提高了突变细胞的回收率，并使其能够在样本中被检测到，这表明在开始RAPD分析之前应考虑DNA修饰，如胁迫诱导的改变和超螺旋现象。
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