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Total synthesis of the cystatin α gene and its expression in E. coli
Author(s) -
Katunuma Nobuhiko,
Yamato Masayuki,
Kominami Eiki,
Ike Yoshimasa
Publication year - 1988
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(88)80238-2
Subject(s) - microbiology and biotechnology , plasmid , gene , pbr322 , oligonucleotide , gene expression , chemistry , sephadex , escherichia coli , biology , restriction enzyme , expression vector , gene product , biochemistry , recombinant dna , enzyme
A gene encoding cystatin α has been chemically synthesized, cloned and expressed in E. coli . The gene of 318 base pairs was assembled by enzymatic ligation of 19 oligonucleotides and cloned into a pBR322‐derived expression plasmid down stream of the tac promoter. The expression product of the synthetic gene has been purified by Sephadex G‐50 column chromatography and shown to have the same properties as those of the authentic protein isolated from rat epidermis.