A synthetic operon containing 14 bovine pancreatic trypsin inhibitor genes is expressed in E. coli.
The EMBO JournalPublished 1 December 1986
B. von Wilcken‐Bergmann, Daniela Tils, J. Sartorius, Ennes A. Auerswald, Wolfgang P. Schröder, Benno Müller‐Hill
Citations36
SJR quartileQ1
SJR score4.82
SNIP1.93
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TL;DR
The cloned BPTI protein is shown to be an efficient inhibitor of human leukocyte elastase and can be reconstituted almost completely after purification.
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.
Keywords
Biochemistry, Genetics and Molecular Biology
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