AI Article Synopsis
- A specific primer pair was created to amplify the 16S rRNA gene from various Bacillus species, resulting in a successful 1114 bp PCR product that did not amplify DNA from 16 other Eubacterial species.
- The amplicons were then analyzed using restriction enzymes, revealing unique patterns for 15 ATCC reference strains among Bacillus, Paenibacillus, and Brevibacillus, as well as identifying three misclassified Bacillus probiotic strains.
- This simplified PCR-ARDRA method offers an effective way to identify important Bacillus species found in environmental sources.
Article Abstract
A group-specific primer pair was designed to amplify the 16S rRNA gene of representative reference strains from environmentally sourced, mesophilic aerobic spore-forming Bacillus taxa. The PCR generated a 1114 bp amplicon but did not do so with DNA extracted from 16 other Eubacterial species. When amplicons were digested with restriction enzymes AluI or TaqI, different profiles containing between 2 and 5 fragments ranging in size from 76 to 804 base pairs were seen with different Bacillus species. This procedure, known otherwise as amplified ribosomal DNA restriction analysis or ARDRA, produced unique and distinguishable patterns to differentiate between 15 ATCC reference strains (10 Bacillus, 3 Paenibacillus and 2 Brevibacillus member species) as well as 3 misidentified Bacillus probiotic strains in a commercial collection. Our simplified PCR-ARDRA protocol provides a facile method for the identification of most environmentally important species of Bacillus.
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| http://dx.doi.org/10.1016/j.mimet.2005.04.021 | DOI Listing |
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