RESUMO
The utility of parallel hybridization of environmental nucleic acids to many oligonucleotides immobilized in a matrix of polyacrylamide gel pads on a glass slide (oligonucleotide microchip) was evaluated. Oligonucleotides complementary to small-subunit rRNA sequences of selected microbial groups, encompassing key genera of nitrifying bacteria, were shown to selectively retain labeled target nucleic acid derived from either DNA or RNA forms of the target sequences. The utility of varying the probe concentration to normalize hybridization signals and the use of multicolor detection for simultaneous quantitation of multiple probe-target populations were demonstrated.
Assuntos
Técnicas Biossensoriais , Microbiologia Ambiental , Sondas de Oligonucleotídeos/genética , Bactérias/genética , Sequência de Bases , Estudos de Avaliação como Assunto , Hibridização de Ácido Nucleico , RNA Bacteriano/genética , RNA Ribossômico/genética , RNA Ribossômico 16S/genéticaRESUMO
Volume 62, no. 6, p. 2157, Table 1: the sequence for probe Nso1225, 5(prm1)-CGCGATTGTATTACGTGTGA-3(prm1), should read 5(prm1)-CGCCATTGTATTACGTGTGA-3(prm1). [This corrects the article on p. 2156 in vol. 62.].
RESUMO
A hierarchical set of five 16S rRNA-targeted oligonucleotide DNA probes for phylogenetically defined groups of autotrophic ammonia- and nitrite-oxidizing bacteria was developed for environmental and determinative studies. Hybridization conditions were established for each probe by using temperature dissociation profiles of target and closely related nontarget organisms to document specificity. Environmental application was demonstrated by quantitative slot blot hybridization and whole-cell hybridization of nitrifying activated sludge and biofilm samples. Results obtained with both techniques suggested the occurrence of novel populations of ammonia oxidizers. In situ hybridization experiments revealed that Nitrobacter and Nitrosomonas species occurred in clusters and frequently were in contact with each other within sludge flocs.
