Synthesis of a (desSer1 Ile29 Leu89) chicken cystatin gene, expression in E. coli as fusion protein and its isolation
- 30 January 1989
- journal article
- Published by Wiley in FEBS Letters
- Vol. 243 (2) , 186-192
- https://doi.org/10.1016/0014-5793(89)80127-9
Abstract
A synthetic gene coding for the cysteine proteinase inhibitor (desSer 1 Ile29 Leu89) chicken cystatin was cloned and expressed in E. coli. The gene was assembled from 12 oligonucleotides and inserted into vector pUC 8. Expression as fusion protein was performed in a temperature-inducible E. coli system. The expression product was synthesized as 20% of total E. coli protein. The fusion protein was purified, the chicken cystatin homologue was split off with CNBr and the N-terminal sequence confirmed up to position 37. The properties of the purified material correspond to those of natural chicken cystatin. The recombinant cystatin variant binds anti-chicken cystatin IgG, is inhibitorily active and displays K i values with papain and with cathepsin B similar to those determined for natural chicken cystatin.Keywords
This publication has 31 references indexed in Scilit:
- Studies on transformation of Escherichia coli with plasmidsPublished by Elsevier ,2006
- Cloning a synthetic gene for human stefin B and its expression in E. coliFEBS Letters, 1988
- Total synthesis of the cystatin α gene and its expression in E. coliFEBS Letters, 1988
- Efficient production of native, biologically active human cystatin C by Escherichia coliFEBS Letters, 1988
- Inhibitorily Active Recombinant Human Stefin B. Gene Synthesis, Expression and Isolation of an Inhibitorily Active MS-2 pol-Stefin B Fusion Protein and Preparation of Des[Met1,22] stefin BBiological Chemistry Hoppe-Seyler, 1988
- Chemical Synthesis of a Gene for Human Stefin A and its Expression inE. coliBiological Chemistry Hoppe-Seyler, 1988
- Synthesis, Cloning and Expression of Recombinant AprotininBiological Chemistry Hoppe-Seyler, 1987
- Protein Inhibitors of Cysteine Proteinases. III. Amino-Acid Sequence of Cystatin from Chicken Egg WhiteHoppe-Seyler´s Zeitschrift Für Physiologische Chemie, 1983
- A Convenient Large-Scale Preparation of High Molecular Weight Kininogen from Human PlasmHoppe-Seyler´s Zeitschrift Für Physiologische Chemie, 1981
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970