DISTRIBUTION OF REPETITIVE DNA-SEQUENCES IN EUBACTERIA AND APPLICATION TO FINGERPRINTING OF BACTERIAL GENOMES
DISTRIBUTION OF REPETITIVE DNA-SEQUENCES IN EUBACTERIA AND APPLICATION TO FINGERPRINTING OF BACTERIAL GENOMES
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DOI：
10.1093/nar/19.24.6823
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发表时间：
1991-12-25
影响因子：
14.9
通讯作者：
LUPSKI, JR
中科院分区：
文献类型：
--
作者：
VERSALOVIC, J;KOEUTH, T;LUPSKI, JR
最近在真细菌中描述了分散的重复DNA序列。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]扩增和狭缝印迹杂交实验中使用了来自不同真细菌物种的基因组DNA的共识寡核苷酸。合成匹配重复基因外回文[REP]元件和肠杆菌重复基因间共有序列[ERIC]的寡核苷酸，并在真细菌基因组DNA的扩增中作为相对PCR引物进行测试。REP和ERIC共有寡核苷酸在PCR扩增后通过琼脂糖凝胶电泳产生清晰可分辨的条带。这些带型提供了不同真细菌种和菌株的明确DNA指纹。REP和ERIC探针都优先与革兰氏阴性肠道细菌和相关物种的基因组DNA杂交。这些重复的DNA元件在各种微生物的基因组中的广泛分布应该能够快速鉴定细菌物种和菌株，并可用于原核基因组的分析。
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.