Use of rpoB and 16S rRNA genes to analyse bacterial diversity of a tropical soil using PCR and DGGE
Use of rpoB and 16S rRNA genes to analyse bacterial diversity of a tropical soil using PCR and DGGE
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DOI：
10.1046/j.1472-765x.2002.01183.x
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发表时间：
2002-10
影响因子：
2.4
通讯作者：
R. Peixoto;H. Coutinho;N. Rumjanek;A. Macrae;A. Rosado
中科院分区：
文献类型：
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作者：
R. Peixoto;H. Coutinho;N. Rumjanek;A. Macrae;A. Rosado
目的：评价rpoB基因在巴西塞拉多土壤DNA PCR - DGGE微生物分析中的应用价值。方法：从土壤中提取DNA，进行rpoB和16S rRNA基因的PCR扩增。PCR产物通过变性梯度凝胶电泳（DGGE）比较基因/群落谱。结果：rpoB DGGE谱比16S rDNA谱包含更少的条带，更容易描绘和分析。通过对两种方法的比较，发现两种方法具有互补性。结论、研究意义及影响：RNA聚合酶β亚基基因rpoB与16S rDNA不同，它是一个单拷贝基因。细菌基因组中16S rRNA基因的多个拷贝使DGGE谱的多样性评估复杂化。使用rpoB基因为基于DGGE的微生物群落分析提供了比常用的16S rRNA基因更好的选择。
Aim: To evaluate the rpoB gene as a biomarker for PCR‐DGGE microbial analyses using soil DNA from the Cerrado, Brazil. Methods: DNA extraction from soil was followed by Polymerase Chain Reaction (PCR) amplification of rpoB and 16S rRNA genes. PCR products were compared by Denaturing Gradient Gel Electrophoresis (DGGE) to compare gene/community profiles. Results: The rpoB DGGE profiles comprised fewer bands than the 16S rDNA profiles and were easier to delineate and therefore to analyse. Comparison of the community profiles revealed that the methods were complementary. Conclusions, Significance and Impact of the Study: The gene for the beta subunit of the RNA polymerase, rpoB, is a single copy gene unlike 16S rDNA. Multiple copies of 16S rRNA genes in bacterial genomes complicate diversity assessments made from DGGE profiles. Using the rpoB gene offers a better alternative to the commonly used 16S rRNA gene for microbial community analyses based on DGGE.