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通过对编码16S rRNA的基因片段进行PCR扩增后进行变性梯度凝胶电泳,确定了半混合湖Saelenvannet中浮游细菌的分布。

Distribution of bacterioplankton in meromictic Lake Saelenvannet, as determined by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA.

作者信息

Ovreås L, Forney L, Daae F L, Torsvik V

机构信息

Department of Microbiology, University of Bergen, Norway.

出版信息

Appl Environ Microbiol. 1997 Sep;63(9):3367-73. doi: 10.1128/aem.63.9.3367-3373.1997.

Abstract

The community structure of bacterioplankton in meromictic Lake Saelenvannet was examined by PCR amplification of the V3 region of 16S rRNA from microbial communities recovered from various depths in the water column. Two different primer sets were used, one for amplification of DNA from the domain Bacteria and another specific for DNA from the domain Archaea. Amplified DNA fragments were resolved by denaturing gradient gel electrophoresis (DGGE), and the resulting profiles were reproducible and specific for the communities from different depths. Bacterial diversity estimated from the number and intensity of specific fragments in DGGE profiles decreased with depth. The reverse was true for the Archaea, with the diversity increasing with depth. Hybridization of DGGE profiles with oligonucleotide probes specific for phylogenetic groups of microorganisms showed the presence of both sulfate-reducing bacteria and methanogens throughout the water column, but they appeared to be most abundant below the chemocline. Several dominant fragments in the DGGE profiles were excised and sequenced. Among the dominant populations were representatives related to Chlorobium phaeovibrioides, chloroplasts from eukaryotic algae, and unidentified Archaea.

摘要

通过对从水柱不同深度采集的微生物群落中16S rRNA的V3区域进行PCR扩增,研究了半咸水湖Saelenvannet中浮游细菌的群落结构。使用了两组不同的引物,一组用于扩增细菌域的DNA,另一组专门用于扩增古菌域的DNA。扩增的DNA片段通过变性梯度凝胶电泳(DGGE)进行分离,所得图谱对于不同深度的群落具有可重复性和特异性。根据DGGE图谱中特定片段的数量和强度估计,细菌多样性随深度降低。古菌的情况则相反,其多样性随深度增加。用针对微生物系统发育组的寡核苷酸探针与DGGE图谱杂交,结果表明水柱中存在硫酸盐还原菌和产甲烷菌,但它们似乎在化学跃层以下最为丰富。切除并测序了DGGE图谱中的几个主要片段。主要种群包括与褐绿菌、真核藻类叶绿体以及未鉴定古菌相关的代表。

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Bacterial population dynamics in a meromictic lake.半咸水湖中的细菌种群动态
Appl Environ Microbiol. 1997 Jun;63(6):2181-8. doi: 10.1128/aem.63.6.2181-2188.1997.

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