Isolation and preliminary characterization of the Streptococcus mutans rpsJ gene.

Abstract

We previously reported that four putative open reading frames were identified in the regions flanking the Streptococcus mutans GS-5 fructosyltransferase gene. For one of these, ORF 4, only a small region had been isolated and the first 30 nucleotides had been sequenced. In order to determine whether this open reading frame is part of an expressed gene, we isolated a DNA fragment containing intact ORF 4 and a portion of the downstream ORF 5 by inverse polymerase chain reaction. A comparison of the deduced amino acid sequences of ORF 4 and ORF 5 with other proteins revealed that the ORF 4 and ORF 5 gene products were highly homologous to ribosomal proteins S10 and L3, respectively, of several bacteria. To identify the precise transcriptional start site for the ORF 4 gene, primer extension analysis was carried out. The results indicated initiation at a G residue with corresponding -10 and -35 regions homologous to the Escherichia coli consensus promoter sequences. These results indicate that the sequences of ORF 4 and ORF 5 are consistent with the structures of ribosomal proteins S10 and L3, respectively, and are present in a ribosomal protein operon.