Abstract
The coding sequence for the human pancreatic ribonuclease (HP-RNase) gene has been obtained from genomic DNA extracted from buccal epithelial cells. In order to direct the expression of the recombinant human pancreatic enzyme to the periplasmic space of E.coli, the bovine pancreatic RNase signal sequence has been fused 5' to the human gene. Initial attempts to express the recombinant enzyme were not successful, consequently site-directed mutagenesis tehniques were used to genetically engineer the HP-RNase gene to enable expression in E.coli. The resultant engineered enzyme shows similar kinetic characteristics to the homologous bovine enzyme.
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