Molecular cloning in Bacillus subtilis of a Bacillus licheniformis gene encoding a thermostable alpha amylase.
A resident-plasmid cloning system developed for Bacillus subtilis has been used to isolate recombinant plasmids carrying DNA from Bacillus licheniformis which confer alpha-amylase activity on alpha-amylase-negative mutants of B. subtilis. These plasmids contain a 3550-bp insert at the EcoRI site of the plasmid pBD64. Subcloning various lengths of the B. licheniformis DNA has localised the gene to a 2550-bp BclI fragment. We present evidence that the cloned fragment codes for a B. licheniformis heat-stable alpha-amylase with a temperature optimum of 93 degrees C. The foreign gene is expressed efficiently in B. subtilis and is stably maintained.
D001407:Bacillus / Q000235:genetics*; D001412:Bacillus subtilis / Q000235:genetics*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D006358:Hot Temperature; D009154:Mutation; D010957:Plasmids; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000235:genetics*
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D000516:alpha-Amylases