Quantification of dissimilatory (bi)sulphite reductase gene expression in Desulfobacterium autotrophicum using real-time RT-PCR.
We developed a real-time RT-PCR method for the quantification of dissimilatory (bi)sulphite reductase (DSR) mRNA in Desulfobacterium autotrophicum cells. The amount of DSR mRNA was determined relative to the amount of 16S rRNA at different growth conditions during transition from exponential to stationary phase: sulphate respiration with lactate, thiosulphate respiration with lactate, sulphate respiration with H2 and pyruvate fermentation. The dsr gene was expressed constitutively, although DSR mRNA content per-cell varied under different growth conditions. The maximum DSR mRNA per-cell content was 2.0 to 4.1-fold higher during sulphate or thiosulphate respiration than during pyruvate fermentation. After transfer of a pyruvate-fermenting culture into sulphate-rich medium, upregulation of the DSR mRNA content was observed. Irrespective of the mode of metabolism the per-cell DSR mRNA content changed significantly during growth (up to 310-fold from the early to the late exponential phase during respiration with thiosulphate). The maximum DSR mRNA per-cell contents correlated with cell-specific sulphate reduction rates for all experiments. Environmental applications for the quantification of DSR mRNA are discussed.
Neretin
Lev N
LN
Biogeochemistry Department, Max Planck Institute for Marine Microbiology, Celsiusstrasse 1, D-28359 Bremen, Germany. lneretin@mpi-bremen.de
Schippers
Axel
A
Pernthaler
Annelie
A
Hamann
Knut
K
Amann
Rudolf
R
Jørgensen
Bo Barker
BB
D003470:Culture Media; D020564:Deltaproteobacteria / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D007773:Lactates / Q000378:metabolism; D010084:Oxidation-Reduction; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000235:genetics* / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D012329:RNA, Bacterial / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013431:Sulfates / Q000378:metabolism; D013885:Thiosulfates / Q000378:metabolism
D003470:Culture Media; D007773:Lactates; D011773:Pyruvates; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012336:RNA, Ribosomal, 16S; D013431:Sulfates; D013885:Thiosulfates; D050862:Oxidoreductases Acting on Sulfur Group Donors