By Northern blot and primer extension analyses it was shown that in Bacillus stearothermophilus the genes infC, rpmI and rplT constitute a single transcriptional unit; the promoter and the transcriptional start-point used in vivo were identified and the half-life of the transcript (1.2 min) was determined. No indication of multiple initiation sites nor of differential stability of different regions of the transcript was found. The results suggest that Escherichia coli and B. stearothermophilus have a different pattern of transcription around the infC gene cluster and that differential translational efficiency within the infC-rpmI-rplT transcriptional unit accounts for the different levels of IF3, L35 and L20 expression. The rare AUU initiation codon is the only strictly conserved element of the several peculiar transcriptional and translational features found in E. coli infC. Upon changing this codon to AUG, we found a ca. 30-fold increased expression of B. stearothermophilus infC, which is similar to that previously found with E. coli infC (i.e. 40-fold), and provided evidence that regulation of infC expression through its rare AUU initiation codon might be a general phenomenon in bacteria.