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...

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Bibliographic Details
Published in:The EMBO journal 1986-12, Vol.5 (12), p.3219-3225
Main Authors: Wilcken‐Bergmann, B., Tils, D., Sartorius, J., Auerswald, E.A., Schröder, W., Müller‐Hill, B.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Aprotinin - genetics
Base Sequence
Biological and medical sciences
Biotechnology
Cattle
Cloning, Molecular
DNA Restriction Enzymes
Escherichia coli
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Gene Amplification
Genes, Synthetic
Genetic engineering
Genetic technics
Methods. Procedures. Technologies
Operon
Plasmids
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary: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.
ISSN:0261-4189
1460-2075
DOI:10.1002/j.1460-2075.1986.tb04632.x