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Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes.

机译:真细菌中重复DNA序列的分布及其在细菌基因组指纹图谱中的应用。

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摘要

Dispersed repetitive DNA sequences have been described recently in eubacteria. To assess the distribution and evolutionary conservation of two distinct prokaryotic repetitive elements, consensus oligonucleotides were used in polymerase chain reaction [PCR] amplification and slot blot hybridization experiments with genomic DNA from diverse eubacterial species. Oligonucleotides matching Repetitive Extragenic Palindromic [REP] elements and Enterobacterial Repetitive Intergenic Consensus [ERIC] sequences were synthesized and tested as opposing PCR primers in the amplification of eubacterial genomic DNA. REP and ERIC consensus oligonucleotides produced clearly resolvable bands by agarose gel electrophoresis following PCR amplification. These band patterns provided unambiguous DNA fingerprints of different eubacterial species and strains. Both REP and ERIC probes hybridized preferentially to genomic DNA from Gram-negative enteric bacteria and related species. Widespread distribution of these repetitive DNA elements in the genomes of various microorganisms should enable rapid identification of bacterial species and strains, and be useful for the analysis of prokaryotic genomes.
机译:最近在真细菌中描述了分散的重复DNA序列。为了评估两个不同的原核重复元件的分布和进化保守性,将共有的寡核苷酸用于聚合酶链反应[PCR]扩增和狭缝印迹杂交实验中,该实验使用了来自多种真细菌的基因组DNA。合成了匹配重复性外基因回文[REP]元件和肠细菌性重复基因间共识[ERIC]序列的寡核苷酸,并作为反向PCR引物进行了测试,以扩增真细菌基因组DNA。 PCR扩增后,通过琼脂糖凝胶电泳,REP和ERIC共有寡核苷酸产生了清晰可分辨的条带。这些条带模式提供了不同真细菌种类和菌株的明确DNA指纹。 REP和ERIC探针均优先与革兰氏阴性肠杆菌和相关物种的基因组DNA杂交。这些重复DNA元素在各种微生物的基因组中的广泛分布应能快速鉴定细菌种类和菌株,并有助于分析原核生物基因组。

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