EXPRESSION OF PSEUDOMONAS-AMYLODERAMOSA ISOAMYLASE GENE IN ESCHERICHIA-COLI USING AN INDUCIBLE T7 PHAGE EXPRESSION SYSTEM

The isoamylase gene (isoA) from Pseudomonas amyloderamosa was amplifie d by polymerase chain reaction (PCR), cloned into the pET-3a vector an d expressed in Escherichia coli. The enzyme from E. coli was immunolog ically identical to that from P. amyloderamonas. The use of isopropyl- beta-D-thiogalactoside (IPTG) for indution of the isoA gene expression resulted in an intracellular insoluble aggregate. Active isoamylase p reparation was obtained by solubilizing and refolding from the inclusi on bodies.