Identification of species belonging to the Bifidobacterium genus by PCR-RFLP analysis of a hsp60 gene fragment
| Autor: | Loredana Baffoni, Verena Stenico, Paola Mattarelli, Monica Marianna Modesto, Francesca Gaggia, Erwin Strahsburger, Bruno Biavati, Diana Di Gioia |
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| Přispěvatelé: | Loredana Baffoni, Verena Stenico, Erwin Strahsburger, Francesca Gaggìa, Diana Di Gioia, Monica Modesto, Paola Mattarelli, Bruno Biavati |
| Rok vydání: | 2013 |
| Předmět: |
DNA
Bacterial Microbiology (medical) bacterial taxonomy Sequence analysis hsp60 Polymerase Chain Reaction Microbiology Actinobacteria PCR-RFLP Phylogenetics Phylogeny Taxonomy Bifidobacterium Genetics biology Methodology Article Bacterial taxonomy Chaperonin 60 Sequence Analysis DNA 16S ribosomal RNA biology.organism_classification Genes Bacterial Bifidobacterium spp Taxonomy (biology) Restriction fragment length polymorphism Polymorphism Restriction Fragment Length |
| Zdroj: | BMC Microbiology |
| ISSN: | 1471-2180 |
| DOI: | 10.1186/1471-2180-13-149 |
| Popis: | BACKGROUND: Bifidobacterium represents one of the largest genus within the Actinobacteria, and includes at present 32 species. These species share a high sequence homology of 16S rDNA and several molecular techniques already applied to discriminate among them give ambiguous results.The slightly higher variability of the hsp60 gene sequences with respect to the 16S rRNA sequences offers better opportunities to design or develop molecular assays, allowing identification and differentiation of closely related species. hsp60 can be considered an excellent additional marker for inferring the taxonomy of the members of Bifidobacterium genus. RESULTS: This work illustrates a simple and cheap molecular tool for the identification of Bifidobacterium species. The hsp60 universal primers were used in a simple PCR procedure for the direct amplification of 590 bp of the hsp60 sequence. The in silico restriction analysis of bifidobacterial hsp60 partial sequences allowed the identification of a single endonuclease (HaeIII) able to provide different PCR-restriction fragment length polymorphism (RFLP) patterns in the Bifidobacterium spp. type strains evaluated. The electrophoretic analyses allowed to confirm the different RFLP patterns. CONCLUSIONS: The developed PCR-RFLP technique resulted in efficient discrimination of the tested species and subspecies and allowed the construction of a dichotomous key in order to differentiate the most widely distributed Bifidobacterium species as well as the subspecies belonging to B. pseudolongum and B. animalis. |
| Databáze: | OpenAIRE |
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