PROFILING OF COMPLEX MICROBIAL-POPULATIONS BY DENATURING GRADIENT GEL-ELECTROPHORESIS ANALYSIS OF POLYMERASE CHAIN REACTION-AMPLIFIED GENES-CODING FOR 16S RIBOSOMAL-RNA
PROFILING OF COMPLEX MICROBIAL-POPULATIONS BY DENATURING GRADIENT GEL-ELECTROPHORESIS ANALYSIS OF POLYMERASE CHAIN REACTION-AMPLIFIED GENES-CODING FOR 16S RIBOSOMAL-RNA
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DOI：
10.1128/aem.59.3.695-700.1993
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发表时间：
1993-03-01
影响因子：
4.4
通讯作者：
UITTERLINDEN, AG
中科院分区：
文献类型：
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作者：
MUYZER, G;DEWAAL, EC;UITTERLINDEN, AG
我们描述了一种分析复杂微生物种群遗传多样性的新分子方法。该技术基于通过变性梯度凝胶电泳（DGGE）对编码16S rRNA且长度相同的基因的聚合酶链反应扩增片段进行分离。对不同微生物群落的DGGE分析表明，在分离图谱中存在多达10条可区分的条带，这些条带很可能来自构成这些种群的许多不同物种，从而产生了这些种群的DGGE图谱。我们表明，可以鉴定出仅占总种群1%的成分。利用一种针对硫酸盐还原菌16S rRNA的V3区域的特异性寡核苷酸探针，通过杂交分析可以鉴定一些微生物种群中的特定DNA片段。对在有氧条件下生长的细菌生物膜的基因组DNA分析表明，尽管硫酸盐还原菌是厌氧的，但它们存在于这种环境中。我们得到的结果表明，该技术将有助于我们理解未被鉴定的微生物种群的遗传多样性。
We describe a new molecular approach to analyzing the genetic diversity of complex microbial populations. This technique is based on the separation of polymerase chain reaction-amplified fragments of genes coding for 16S rRNA, all the same length, by denaturing gradient gel electrophoresis (DGGE). DGGE analysis of different microbial communities demonstrated the presence of up to 10 distinguishable bands in the separation pattern, which were most likely derived from as many different species constituting these populations, and thereby generated a DGGE profile of the populations. We showed that it is possible to identify constituents which represent only 1% of the total population. With an oligonucleotide probe specific for the V3 region of 16S rRNA of sulfate-reducing bacteria, particular DNA fragments from some of the microbial populations could be identified by hybridization analysis. Analysis of the genomic DNA from a bacterial biofilm grown under aerobic conditions suggests that sulfate-reducing bacteria, despite their anaerobicity, were present in this environment. The results we obtained demonstrate that this technique will contribute to our understanding of the genetic diversity of uncharacterized microbial populations.