A synthetic operon containing 14 bovine pancreatic trypsin inhibitor genes is expressed in E. coli.
A synthetic gene encoding the protein sequence of mature bovine pancreatic trypsin inhibitor (BPTI) has been cloned into a novel E. coli expression vector. After in vitro gene amplification by successive DNA duplications, more than 600 000 mostly inactive inhibitor molecules may be recovered from a single cell. After purification the inhibitory activity can be reconstituted almost completely. The specificity of BPTI for trypsin is abolished by a single amino acid exchange from lysine to isoleucine at position 15. The altered protein is shown to be an efficient inhibitor of human leukocyte elastase.
D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005784:Gene Amplification; D005813:Genes, Synthetic; D009876:Operon; D010957:Plasmids
D007611:Aprotinin; D004262:DNA Restriction Enzymes