Ribosomal RNA (rrn) operons in Streptococcus mutans and nucleotide sequence of tRNA(Pro) gene associated with rrnB

Using a cloned 0.5-kb probe containing an internal fragment of 23S ribosomal RNA from the rrnB operon of Streptococcus mutans, we analyzed various endonuclease digests of the chromosomal DNA isolated from human-derived strains of mutans streptococci. Thus far, the examined S. mutans strains exhibited five ribosomal operons. Here, we describe a ribotyping technique for S. mutans based on restriction and Southern blot analyses with the biotin-labeled homologous probe and chemiluminescence detection. We cloned and sequenced a unique gene encoding tRNA(Pro) downstream from 23S rRNA gene at the 3' end of the operon. Primers designed to the 3' end of the rrnB operon PCR-amplified a 2.3-kb DNA fragment in all tested strains. Restriction fragment length polymorphism analysis of the amplicon revealed a diversity of the single locus among S. mutans isolates, thus establishing a potential use of the technique for the molecular epidemiology of mutans streptococci.