DsrB gene-based DGGE for community and diversity surveys of sulfate-reducing bacteria.
A denaturing gradient gel electrophoresis (DGGE) method was developed to assess the diversity of dsrB (dissimilatory sulfite reductase beta-subunit)-genes in sulfate-reducing communities. For this purpose a PCR primer pair was optimized for the amplification of a approximately 350 bp dsrB gene fragment that after DGGE gel electrophoresis enabled us to discriminate between dsrB genes of different SRB-subgroups,-genera and -species. The dsrB-DGGE method revealed considerable genetic diversity when applied to DNA extracts obtained from aquifer samples that were derived from monitoring wells of an in situ metal precipitation (ISMP) pilot project conducted at the site of a non-ferrous industry or from environmental heavy metal contaminated samples. The sequences of the excised and sequenced DGGE bands represented dsrB genes of different SRB-subgroups,-genera and -species, thus confirming the broad applicability of the PCR primer pair. Linking the results of the physico-chemical follow-up of the field and lab experiments to the dsrB-DGGE data will provide a better understanding of the contribution of the SRB populations to the ongoing ISMP processes.
Geets
Joke
J
Limburg University Centrum, Department of Chemistry, Biology and Geology, Environmental Biology Group, Universitaire Campus, B-3590 Diepenbeek, Belgium.
Borremans
Brigitte
B
Diels
Ludo
L
Springael
Dirk
D
Vangronsveld
Jaco
J
van der Lelie
Daniel
D
Vanbroekhoven
Karolien
K
D001483:Base Sequence; D001673:Biodegradation, Environmental; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D014644:Genetic Variation; D050885:Hydrogensulfite Reductase / Q000235:genetics*; D019216:Metals, Heavy / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D016999:Sulfur-Reducing Bacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D014874:Water Pollutants, Chemical / Q000378:metabolism
D017931:DNA Primers; D004269:DNA, Bacterial; D019216:Metals, Heavy; D014874:Water Pollutants, Chemical; D050885:Hydrogensulfite Reductase