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. 1986 Dec 1;5(12):3219–3225. doi: 10.1002/j.1460-2075.1986.tb04632.x

A synthetic operon containing 14 bovine pancreatic trypsin inhibitor genes is expressed in E. coli.

B von Wilcken-Bergmann, D Tils, J Sartorius, E A Auerswald, W Schröder, B Müller-Hill
PMCID: PMC1167315  PMID: 2434325

Abstract

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.

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