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New primers for the class Actinobacteria: application to marine and terrestrial environments.

Stach, James E.M., Maldonado, Luis A., Ward, Alan C., Goodfellow, Michael, Bull, Alan T. (2003) New primers for the class Actinobacteria: application to marine and terrestrial environments. Environmental Microbiology, 5 (10). pp. 828-841. ISSN 1462-2912. E-ISSN 1462-2920. (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:86)

Language: English

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In this study, we redesigned and evaluated primers for the class Actinobacteria. In silico testing showed that the primers had a perfect match with 82% of genera in the class Actinobacteria, representing a 26-213% improvement over previously reported primers. Only 4% of genera that displayed mismatches did so in the terminal three bases of the 3' end, which is most critical for polymerase chain reaction success. The primers, designated S-C-Act-0235-a-S-20 and S-C-Act-0878-a-A-19, amplified an approximately 640 bp stretch of the 16S rRNA gene from all actinobacteria tested (except Rubrobacter radiotolerans) up to an annealing temperature of 72 degrees C. An Actinobacteria Amplification Resource ( was generated to provide a visual guide to aid the amplification of actinobacterial 16S rDNA. Application of the primers to DNA extracted from marine and terrestrial samples revealed the presence of actinobacteria that have not been described previously. The use of 16S rDNA similarity and DNA-DNA pairing correlations showed that almost every actinomycete clone represented either a new species or a novel genus. The results of this study reinforce the proposition that current culture-based techniques drastically underestimate the diversity of Actinobacteria in the environment and highlight the need to evaluate taxon-specific primers regularly in line with improvements in databases holding 16S rDNA sequences.

Item Type: Article
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Susan Davies
Date Deposited: 19 Dec 2007 17:55 UTC
Last Modified: 16 Nov 2021 09:39 UTC
Resource URI: (The current URI for this page, for reference purposes)
Bull, Alan T.:
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