Author/Editor		Auerswald, EA; Genenger, G; Assfalg-Machleidt, I; Kos, J; Bode, W
Title		Synthesis of a (desSer1 Ile29 Leu89) chicken cystatin gene, expression in E. coli as fusion protein and its isolation
Type		članek
Source		FEBS Lett
Vol. and No.		Letnik 243, št. 2
Publication year		1989
Volume		str. 186-92
Language		eng
Abstract		A synthetic gene coding for the cysteine proteinase inhibitor (desSer1 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 percent 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 Ki values with papain and with cathepsin B similar to those determined for natural chicken cystatin.
Descriptors		ESCHERICHIA COLI GENES, SYNTHETIC PROTEINS AMINO ACID SEQUENCE BASE SEQUENCE CHICKENS CLONING, MOLECULAR GENETIC VECTORS MOLECULAR SEQUENCE DATA PROTEINS RECOMBINANT FUSION PROTEINS