Versalovic J, Koeuth T, Lupski J R
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Nucleic Acids Res. 1991 Dec 25;19(24):6823-31. doi: 10.1093/nar/19.24.6823.
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.
分散的重复DNA序列最近在真细菌中被发现。为了评估两种不同的原核重复元件的分布和进化保守性,在聚合酶链反应(PCR)扩增以及与来自不同真细菌物种的基因组DNA进行狭缝印迹杂交实验中使用了共有寡核苷酸。合成了与重复基因外回文(REP)元件和肠杆菌重复基因间共有序列(ERIC)匹配的寡核苷酸,并作为反向PCR引物用于真细菌基因组DNA的扩增测试。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物种和菌株明确无误的DNA指纹图谱。REP和ERIC探针都优先与革兰氏阴性肠道细菌及相关物种的基因组DNA杂交。这些重复DNA元件在各种微生物基因组中的广泛分布应能实现细菌物种和菌株的快速鉴定,并有助于原核基因组的分析。