Ratios between Alpha-, Beta- and Gamma-proteobacteria in tap water determined by the ProteoQuant assay.
OBJECTIVE
Proteobacteria are widespread on earth. Recently, it has been discovered that a diverse repertoire of proteobacteria are also dominant in tap water. It is therefore important to use high-throughput monitoring tools for tap water. Here, the high-throughput assay ProteoQuant was developed to quantify the main proteobacterial phyla in tap water.


RESULTS
The principle of ProteoQuant is proteobacterial-selective 23S rRNA gene PCR amplification, with multiple competitive TaqMan probes for quantifying the phyla Alpha-, Beta- and Gamma-proteobacteria. The ProteoQuant assay was evaluated, analysing both designed proteobacterial mixes and rRNA gene clone libraries from tap water. These evaluations showed a good coverage and accuracy of the ProteoQuant assay.


CONCLUSIONS
Large-scale tap water screening using ProteoQuant revealed a dominance of Beta-proteobacteria and a potential interaction between Alpha- and Beta-proteobacteria. Gamma-proteobacteria, on the other hand, seemed independent of the two other phyla.


CONCLUSIONS
The ProteoQuant assay will potentially be important for future understanding of the ecological forces shaping the tap water microbiota.
Rudi
K
K
Hedmark University College, Hamar, Norway. knut.rudi@hihm.no
Berg
F
F
Gaustad
E
E
Tannes
T
T
Vatn
M
M
D020561:Alphaproteobacteria / Q000235:genetics / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D020562:Betaproteobacteria / Q000235:genetics / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis; D020563:Gammaproteobacteria / Q000235:genetics / Q000302:isolation & purification*; D020459:Genes, rRNA; D012329:RNA, Bacterial / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature; D014871:Water Microbiology
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S