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利用重复序列聚合酶链反应指纹图谱分析偶蹄目动物源大肠杆菌的异质性。

Heterogeneity of Escherichia coli derived from artiodactyla animals analyzed with the use of rep-PCR fingerprinting.

作者信息

Baldy-Chudzik K, Niedbach J, Stosik M

机构信息

Department of Microbiology and Genetics, Institute of Biotechnology and Environmental Protection, University of Zielona Góra, 65-561 Zielona Góra, Poland.

出版信息

Folia Microbiol (Praha). 2003;48(2):162-7. doi: 10.1007/BF02930949.

Abstract

Genetic polymorphism of 83 isolates of E. coli, derived from 4 species of artiodactyla animals living in a relatively close contact on the grounds of a theme park ZOO Safarii Swierkocin (Poland) was determined using the rep-PCR fingerprinting method, which utilizes oligonucleotide primers matching interspersed repetitive DNA sequences in PCR reaction to yield DNA fingerprints of individual bacterial isolates based on repetitive extragenic palindrome (REP) primers. The fingerprint patterns demonstrated the essential polymorphism of distribution of REP sequences in genomes of the examined isolates. The arithmetic averages clustering algorithm (UPGMA) statistical analysis of fingerprints with the use of the Jaccard similarity coefficient differentiated E. coli isolates into three similarity groups containing various numbers of isolates. The groups comprised isolates derived from two, three and four species of the source animals. The isolates derived from each source segregated in the dendrogram in a different way, both within the similarity groups and among them, indicating an individual repertoire of E. coli in the examined species of animals. The similarity relations among E. coli derived from the same source, illustrated in a dendrogram with a number of subclusters of a low mutual similarity (< or = 20%), indicated an essential interstrain differentiation in terms of the distribution of REP sequences. Our results confirmed the hypothesis of the oligoclonal characters of populations obtained from particular sources. The rep-PCR fingerprinting method with REP primers is simple and highly differentiating and can be recommended for use in explorations of large groups of animals and monitoring the variability of strains.

摘要

利用重复聚合酶链反应(rep-PCR)指纹图谱法,对来自波兰斯维尔科钦动物园Safarii主题公园中4种有蹄类动物的83株大肠杆菌分离株的遗传多态性进行了测定。该方法在聚合酶链反应中使用与散布的重复DNA序列匹配的寡核苷酸引物,基于重复外显子回文序列(REP)引物产生单个细菌分离株的DNA指纹图谱。指纹图谱显示了所检测分离株基因组中REP序列分布的基本多态性。使用Jaccard相似系数对指纹图谱进行算术平均聚类算法(UPGMA)统计分析,将大肠杆菌分离株分为三个相似性组,每组包含不同数量的分离株。这些组包括来自两种、三种和四种源动物的分离株。来自每个源的分离株在聚类图中的分离方式不同,无论是在相似性组内还是组间,这表明在所检测的动物物种中大肠杆菌具有个体特征。来自同一源的大肠杆菌之间的相似性关系在聚类图中表现为多个相互相似性较低(≤20%)的亚群,这表明在REP序列分布方面存在显著的菌株间差异。我们的结果证实了从特定来源获得的群体具有寡克隆特征的假设。使用REP引物的rep-PCR指纹图谱法简单且具有高度区分性,可推荐用于对大量动物群体的探索以及监测菌株的变异性。

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