4.3 Article

Use of rpo B and 16S rRNA genes to analyse bacterial diversity of a tropical soil using PCR and DGGE

Journal

LETTERS IN APPLIED MICROBIOLOGY
Volume 35, Issue 4, Pages 316-320

Publisher

BLACKWELL PUBLISHING LTD
DOI: 10.1046/j.1472-765X.2002.01183.x

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Aim: To evaluate the rpo B 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 rpo B and 16S rRNA genes. PCR products were compared by Denaturing Gradient Gel Electrophoresis (DGGE) to compare gene/community profiles. Results: The rpo B 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, rpo B, 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 rpo B gene offers a better alternative to the commonly used 16S rRNA gene for microbial community analyses based on DGGE.

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