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希瓦氏菌属特异性16S rRNA引物的设计及其在贫营养湿地希瓦氏菌属分析中的应用

Design of Shewanella-specific 16S rRNA primers and application to analysis of Shewanella in a minerotrophic wetland.

作者信息

Todorova Svetoslava G, Costello Andria M

机构信息

Department of Civil and Environmental Engineering, 151 Link Hall, Syracuse University, Syracuse, NY, USA.

出版信息

Environ Microbiol. 2006 Mar;8(3):426-32. doi: 10.1111/j.1462-2920.2005.00908.x.

Abstract

In this study, an existing probe was used as a polymerase chain reaction (PCR) primer to study iron-reducing members of the genus Shewanella in a minerotrophic wetland where iron reduction had previously been implicated. The probe was found to be non-specific and a new set of PCR primers were developed that were specific for Shewanella. These primers were used to analyse the wetland iron-reducing communities by characterizing 16S rRNA genes amplified from DNA extracted from peat. Polymerase chain reaction clone libraries were screened using restriction fragment length polymorphism and diagnostic operational taxonomic units for Shewanella species were identified. A statistical method was used to determine the coverage of the clone libraries, which was found to be between 83% and 97%. The dominant species in the wetland samples at two geochemically distinct zones were phylogenetically related to the iron-reducing microorganism Shewanella oneidensis.

摘要

在本研究中,一种现有的探针被用作聚合酶链反应(PCR)引物,以研究在先前已涉及铁还原的矿质营养湿地中希瓦氏菌属的铁还原成员。发现该探针具有非特异性,因此开发了一组新的针对希瓦氏菌属的PCR引物。这些引物用于通过表征从泥炭中提取的DNA扩增的16S rRNA基因来分析湿地铁还原群落。使用限制性片段长度多态性筛选聚合酶链反应克隆文库,并鉴定希瓦氏菌属物种的诊断操作分类单元。使用一种统计方法来确定克隆文库的覆盖率,发现其在83%至97%之间。在两个地球化学特征不同的区域的湿地样本中的优势物种在系统发育上与铁还原微生物奥奈达希瓦氏菌相关。

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