Division of Biochemistry, Department of Biological Sciences, The University of Calgary, Calgary, Alberta T2N 1N4, and Department of Microbiology, University of Alberta, Edmonton, Alberta T6G 2E9, Canada.
Appl Environ Microbiol. 1991 Nov;57(11):3070-8. doi: 10.1128/aem.57.11.3070-3078.1991.
A novel method for the identification of bacteria in environmental samples by DNA hybridization is presented. It is based on the fact that, even within a genus, the genomes of different bacteria may have little overall sequence homology. This allows the use of the labeled genomic DNA of a given bacterium (referred to as a "standard") to probe for its presence and that of bacteria with highly homologous genomes in total DNA obtained from an environmental sample. Alternatively, total DNA extracted from the sample can be labeled and used to probe filters on which denatured chromosomal DNA from relevant bacterial standards has been spotted. The latter technique is referred to as reverse sample genome probing, since it is the reverse of the usual practice of deriving probes from reference bacteria for analyzing a DNA sample. Reverse sample genome probing allows identification of bacteria in a sample in a single step once a master filter with suitable standards has been developed. Application of reverse sample genome probing to the identification of sulfate-reducing bacteria in 31 samples obtained primarily from oil fields in the province of Alberta has indicated that there are at least 20 genotypically different sulfate-reducing bacteria in these samples.
本文提出了一种通过 DNA 杂交鉴定环境样本中细菌的新方法。其依据是，即使在一个属内，不同细菌的基因组之间可能也只有很少的整体序列同源性。这使得我们可以使用特定细菌的标记基因组 DNA（称为“标准”）来探测其存在以及在从环境样本中获得的总 DNA 中具有高度同源基因组的细菌的存在。或者，也可以对从样本中提取的总 DNA 进行标记，并用于探测在其上点样有相关细菌标准的变性染色体 DNA 的滤器。后一种技术被称为反向样本基因组探测，因为它与通常的做法相反，即从参考细菌中衍生探针来分析 DNA 样本。一旦开发出带有合适标准的主滤器，反向样本基因组探测就可以在单个步骤中鉴定样本中的细菌。将反向样本基因组探测应用于鉴定主要从艾伯塔省油田获得的 31 个样本中的硫酸盐还原菌，表明这些样本中至少存在 20 种不同基因型的硫酸盐还原菌。
