[Cloning of the gene for extracellular Bacillus circulans RNAase]

The gene for extracellular low molecular weight ribonuclease of Bacillus circulans BCF 247 was cloned. The strain was isolated from permafrost deposits of the Kolyma lowland. The gene for the ribonuclease from Bacillus intermedius (binase) was used as a specific probe. The cloning succeeded only in the E. coli strain producing the inhibitor of ribonuclease form Bacillus amyloliquefaciens. Selected clones secreted the active ribonuclease into the growth media. Deletion derivatives of the parental recombinant plasmid were constructed. The smallest DNA fragment which enclosed a functional ribonuclease gene in E. coli was determined to be 0.6 kb in length.