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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Sets of EcoRI fragments containing ribosomal RNA sequences are conserved among different strains of Listeria monocytogenes.
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Sets of EcoRI fragments containing ribosomal RNA sequences are conserved among different strains of Listeria monocytogenes.

机译：含有核糖体RNA序列的EcoRI片段组在单核细胞增生李斯特菌的不同菌株之间是保守的。

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摘要

To classify Listeria monocytogenes using taxonomic characters derived from the rRNA operons and their flanking sequences, we studied a sample of 1346 strains within the taxon. DNA from each strain was digested with a restriction endonuclease, EcoRI. The fragments were separated by gel electrophoresis, immobilized on a membrane, and hybridized with a labeled rRNA operon from Escherichia coli. The pattern of bands, positions, and intensities of hybridized fragments were electronically captured. Software was used to normalize the band positions relative to standards, scale the signal intensity, and reduce the background so that each strain was reproducibly represented in a data base as a pattern. With these methods, L. monocytogenes was resolved into 50 pattern types differing in the length of at least one polymorphic fragment. Pattern types representing multiple strains were recorded as the mathematical average of the strain patterns. Pattern types were arranged by size polymorphisms of assigned rRNA regions into subsets, which revealed the branching genetic structure of the species. Subtracting the polymorphic variants of a specific assigned region from the pattern types and averaging the types within each subset resulted in reduced sets of conserved fragments that could be used to recognize strains of the species. Pattern types and reduced sets of conserved fragments were conserved among different strains of L. monocytogenes but were not observed in total among strains of other species.
机译：为了使用源自rRNA操纵子及其侧翼序列的分类学特征对单核细胞增生李斯特菌进行分类，我们研究了分类单元内1346株菌株的样本。用限制性核酸内切酶EcoRI消化每个菌株的DNA。通过凝胶电泳分离片段，固定在膜上，并与来自大肠杆菌的标记的rRNA操纵子杂交。电子捕获杂交片段的带，位置和强度的模式。使用软件对相对于标准的谱带位置进行归一化，缩放信号强度并减少背景，以使每种菌株都可再现地在数据库中以图案表示。用这些方法，将单核细胞增生李斯特氏菌分解为50种模式类型，这些模式类型的至少一个多态性片段的长度不同。代表多个应变的模式类型被记录为应变模式的数学平均值。模式类型通过分配的rRNA区域的大小多态性排列成子集，从而揭示了该物种的分支遗传结构。从模式类型中减去特定指定区域的多态性变体，然后对每个子集中的类型进行平均，可以减少可用于识别物种菌株的保守片段集。在不同的单核细胞增生李斯特氏菌菌株中，模式类型和减少的保守片段集是保守的，但在其他物种的菌株中却没有观察到。

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