Reverse Transcription of 16S rRNA To Monitor Ribosome-Synthesizing Bacterial Populations in the Environment

摘要: ABSTRACT Identification and quantification of phylogenetically defined bacterial populations in the environment are often performed using molecular tools targeting 16S rRNA. Fluorescence situ hybridization has been used to monitor expression processing rRNA by 3′ tail precursor To expand this approach, we employed reverse transcription total RNA primer S-D-Bact-0338-a-A-18. Length heterogeneity detected slab gel analysis, denaturing high-performance liquid chromatography (DHPLC) was differentiate 5′ from mature rRNA, relative abundance compared shown be a sensitive indicator physiologic state Acinetobacter calcoaceticus ATCC 23055T. Our results demonstrate that is reliable method with detection limit 10 ng single-stranded DNA. The assay also among levels mixed pure cultures, as well examine response activated sludge culture exposed fresh growth medium antibiotic chloramphenicol. study novel simultaneously measures pools for an engineered environment. Furthermore, collection products derived samples DHPLC approach enabled identification active genera. Comparison clone library indicated more bacteria environmental samples.