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Nucleotide Sequencing and Transcriptional Analysis of Two Tandem Genes Encoding Glucosyltransferase (Water‐Soluble‐Glucan Synthetase) in Streptococcus cricetus HS‐6

Two tandem genes encoding glucosyltransferase synthesizing water‐soluble glucan (GTF‐S) were cloned from the lambda gene library of Streptococcus cricetus HS‐6 (serotype a) using anti‐GTF‐S antibody, and the nucleotide sequences were analyzed. The two genes (ORF1 and ORF2) were identified as streptococcal glucosyltransferases based on the following evidence: [1] the deduced amino acid sequences of their products have an active site for catalytic action and C‐terminal repeated units for dextran binding, and [2] a homology search revealed that the ORF1 and ORF2 products are homologous to the GtfS protein (77.4%) of S. downei Mfe28 and GtfT protein (83.8%) of S. sobrinus OMZ176, respectively, which are both known to have GTF‐S activity. Therefore, ORF1 and ORF2 might be designated gtfS and gtfT of S. cricetus, respectively. A Northern blotting and RNase protection assay suggested that the gtfS and gtfT of S. cricetus are transcribed as a single bicistronic mRNA as well as separate monocistronic mRNAs. Primer extension analysis indicated multiple transcriptional start points for each gene.