[Use of the thermoinducible temperate phage Phi 105cts139 for cloning in Bacillus subtilis]

The gene for alpha-amylase from Bacillus amyloliquefaciens having a foreign promoter providing gene expression in logarithmic growth phase and the cat gene of plasmid pC194 (AC fragment) were inserted into thermoinducible prophage phi 105 cts139. Possibility of amylolytic activity enhancement was studied after thermoinduction. When AC fragment and random PstI restricts of phage DNA were ligated and used to transform Bacillus subtilis 1A289 (phi 105 cts139) the Amy+ CmR transformants were obtained having the different levels of increased amylolytic activity (maximum--26 fold). Numerous phages without insert found in induced lysates suggest that insertions were unstable and (or) the clones were double lysogens for hybrid and original type phages. Stable insertion of AC fragment replacing the PstI-H-fragment of phage DNA revealed that all Amy+ CmR transformants were double lysogens. Inducibility depended on the insertion orientation.