Isolation and characterization of a 2.2‐kb operon preceding the α‐amylase gene of Bacillus amyloliquefaciens

A DNA region of 2.8 X 10(3) base pairs (2.8 kb) upstream of the Bacillus amyloliquefaciens alpha-amylase gene has been isolated. This DNA gave rise to a 2.2-kb transcript. The 3' end of the transcript was mapped with S1 nuclease and shown to terminate 49 base pairs upstream of the -35 region of the alpha-amylase promoter. In B. subtilis minicells this 2.2-kb transcript coded for three different polypeptides, thus indicating a polycistronic operon-type structure. The location and the order of the polypeptides were established using DNA deletions. The joining of the 2.2-kb operon to the downstream alpha-amylase gene in the plasmid pUB110 did not have any significant effect on the level of expression of the alpha-amylase.