Novel approach to quantitative detection of specific rRNA in a microbial community, using catalytic DNA.
We developed a novel method for the quantitative detection of the 16S rRNA of a specific bacterial species in the microbial community by using deoxyribozyme (DNAzyme), which possesses the catalytic function to cleave RNA in a sequence-specific manner. A mixture of heterogeneous 16S rRNA containing the target 16S rRNA was incubated with a species-specific DNAzyme. The cleaved target 16S rRNA was separated from the intact 16S rRNA by electrophoresis, and then their amounts were compared for the quantitative detection of target 16S rRNA. This method was used to determine the abundance of the 16S rRNA of a filamentous bacterium, Sphaerotilus natans, in activated sludge, which is a microbial mixture used in wastewater treatment systems. The result indicated that this DNAzyme-based approach would be applicable to actual microbial communities.
Suenaga
Hikaru
H
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8566, Japan. suenaga-hikaru@aist.go.jp
Liu
Rui
R
Shiramasa
Yuko
Y
Kanagawa
Takahiro
T
D001483:Base Sequence; D021881:DNA, Catalytic / Q000378:metabolism*; D004586:Electrophoresis; D008969:Molecular Sequence Data; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000235:genetics / Q000378:metabolism*; D012722:Sewage / Q000382:microbiology*; D013045:Species Specificity; D042744:Sphaerotilus / Q000235:genetics* / Q000378:metabolism; D014865:Waste Disposal, Fluid / Q000379:methods
D021881:DNA, Catalytic; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012722:Sewage