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ARDRA studies of the ribosomal RNA operon within the Desulfovibrio desulfuricans strains.
1
Department of Biopharmacy, Medical University of Silesia, Poland
2
Department of Biotechnology and Genetic Engineering, Medical University of Silesia, Poland
Corresponding author
Joanna Szczerba
Department of Biopharmacy Medical University of Silesia 41-200 Sosnowiec, Poland Narcyzów 1St. tel.: (+48 32) 364 10 61
Department of Biopharmacy Medical University of Silesia 41-200 Sosnowiec, Poland Narcyzów 1St. tel.: (+48 32) 364 10 61
Ann. Acad. Med. Siles. 2009;63:7-19
KEYWORDS
ABSTRACT
BACKGROUND:
Desulfovibrio desulfuricans belong to the heterogeneous group of anaerobic, sulphate-reducing bacteria (SRB), widely distributed in various environments. As a result of dissimilatory sulphate reduction, they release hydrogen sulphide (H2S), which has a cytotoxic effect in human and animal organisms. It has been shown by many authors, that Desulfovibrio was the genus predominating in patients with ulcerative colitis. Some of these bacteria can act as opportunistic pathogens associated with primary bacteremia and abdominal infections such as abscesses.
MATERIAL AND METHODS:
Fifteen (soil and intestinal) strains of Desulfovibrio desulfuricans species were cultured in modified sulphate-free Postgate’s liquid medium with pyruvate for 10 days. Bacterial DNA was extracted by using a commercially available kit and DNA was used as a template for amplification of the full-length 16S, 23S rDNA and the intergenic spacer region. Digested with restriction enzymes (AluI, EcoRI, HaeIII, HindIII, HinfI, MboI and PstI) PCR amplicons were resolved by electrophoresis on 2% agarose gels.
RESULTS:
Digestion of rrn operon of Desulfovibrio desulfuricans by seven restriction enzymes allowed to obtain the characteristic restriction profiles for all 15 investigated strains. The results allow us to suggest three of used enzymes: HinfI, AluI and HaeIII as a useful for confirmation of the similarity within of rrn operon of isolates belonging to this species. onsidering the restriction profiles received with HindIII, and EcoR1 enzymes it seems that their application is insufficient, but PstI enzyme is not acceptable for the analysis of rrn operon of these bacteria.
CONCLUSIONS:
The obtained data have shown that ARDRA can be used for establishment of phylogenetic relations among isolates of Desulfovibrio desulfuricans species, providing the appropriate restriction enzyme is used.
Desulfovibrio desulfuricans belong to the heterogeneous group of anaerobic, sulphate-reducing bacteria (SRB), widely distributed in various environments. As a result of dissimilatory sulphate reduction, they release hydrogen sulphide (H2S), which has a cytotoxic effect in human and animal organisms. It has been shown by many authors, that Desulfovibrio was the genus predominating in patients with ulcerative colitis. Some of these bacteria can act as opportunistic pathogens associated with primary bacteremia and abdominal infections such as abscesses.
MATERIAL AND METHODS:
Fifteen (soil and intestinal) strains of Desulfovibrio desulfuricans species were cultured in modified sulphate-free Postgate’s liquid medium with pyruvate for 10 days. Bacterial DNA was extracted by using a commercially available kit and DNA was used as a template for amplification of the full-length 16S, 23S rDNA and the intergenic spacer region. Digested with restriction enzymes (AluI, EcoRI, HaeIII, HindIII, HinfI, MboI and PstI) PCR amplicons were resolved by electrophoresis on 2% agarose gels.
RESULTS:
Digestion of rrn operon of Desulfovibrio desulfuricans by seven restriction enzymes allowed to obtain the characteristic restriction profiles for all 15 investigated strains. The results allow us to suggest three of used enzymes: HinfI, AluI and HaeIII as a useful for confirmation of the similarity within of rrn operon of isolates belonging to this species. onsidering the restriction profiles received with HindIII, and EcoR1 enzymes it seems that their application is insufficient, but PstI enzyme is not acceptable for the analysis of rrn operon of these bacteria.
CONCLUSIONS:
The obtained data have shown that ARDRA can be used for establishment of phylogenetic relations among isolates of Desulfovibrio desulfuricans species, providing the appropriate restriction enzyme is used.
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