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Molecular cloning in Bacillus subtilis of a Bacillus licheniformis gene encoding a thermostable alpha amylase |
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| Authors: | S A Ortlepp J F Ollington D J McConnell |
| Institution: | Department of Bioiogy Faculty of Science Kyushu University 33. Fukuoka 812 Japan Tel. 092-641-1101 |
| Abstract: | 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. |
| Keywords: | Recombinant DNA pBR322-based plasmid HAT selection shuttle vector amp ampicillin BPV bovine papilloma virus EtBr ethidium bromide HAT medium see MATERIALS AND METHODS section a HSV kb kilobase pairs SDS sodium dodecyl sulfate thymidine kinase |
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