MONITORING THE ENRICHMENT AND ISOLATION OF SULFATE-REDUCING BACTERIA BY USING OLIGONUCLEOTIDE HYBRIDIZATION PROBES DESIGNED FROM ENVIRONMENTALLY DERIVED 16S RIBOSOMAL-RNA SEQUENCES
MONITORING THE ENRICHMENT AND ISOLATION OF SULFATE-REDUCING BACTERIA BY USING OLIGONUCLEOTIDE HYBRIDIZATION PROBES DESIGNED FROM ENVIRONMENTALLY DERIVED 16S RIBOSOMAL-RNA SEQUENCES
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DOI：
10.1128/aem.59.3.682-686.1993
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发表时间：
1993-03-01
影响因子：
4.4
通讯作者：
STAHL, DA
中科院分区：
文献类型：
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作者：
KANE, MD;POULSEN, LK;STAHL, DA
一种荧光标记版本的群体特异性寡核苷酸杂交探针用于监测硫酸盐还原细菌从多物种厌氧生物反应器的富集和分离。通过扩增和测序部分16S rRNA序列，该生物最初被鉴定为与普通脱硫弧菌（Desulfovibrio vulgaris）系统发育相关的分子分离物。该序列依次被用于设计特定种群的探针。用于微生物富集和分离的厌氧培养基是基于其近亲的生理特性，通过序列比较确定。在检测的30株分离株中，只有3株与探针杂交。这三个分离株的16S rRNA序列几乎完全一致(i)与探针靶位没有错配，（ii）与扩增的约500个核苷酸的部分序列完全相同，并且在所有其他位置彼此相同，（iii）与D. vulgaris相似度为93.9%。此外，进一步研究的一个分离株（菌株PT-2）具有与D. vulgaris相当的底物特异性。这些结果证实，从环境样本中扩增16S rRNA序列的聚合酶链反应是准确的，并且还可以提供系统发育信息，从中可以推断种群生理的各个方面。
A fluorescently labeled version of a population-specific oligonucleotide hybridization probe was used to monitor the enrichment and isolation of a sulfate-reducing bacterium from a multispecies anaerobic bioreactor. The organism was originally identified as a molecular isolate that was phylogenctically related to Desulfovibrio vulgaris by amplification and sequencing of part of its 16S rRNA sequence. The sequence, in turn, was used to design a population-specific probe. The anaerobic medium used for the organism's enrichment and isolation was based on the physiological properties of the its closest relatives as identified by sequence comparisons. Of 30 isolates examined, only 3 hybridized with the probe. Nearly complete 16S rRNA sequences determined for each of these three isolates (i) had no mismatches with the probe target site, (ii) were identical to the amplified partial sequence of about 500 nucleotides and to one another in all other positions, and (iii) were 93.9% similar to that of D. vulgaris. In addition, one isolate chosen for further study (strain PT-2) had a substrate specificity comparable to that of D. vulgaris. These results confirmed that polymerase chain reaction amplification of 16S rRNA sequences from environmental samples can be accurate and can also provide phylogenetic information from which aspects of a population's physiology can be inferred.