Total synthesis of the cystatin alpha gene and its expression in E. coli.
A gene encoding cystatin alpha 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.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D015853:Cysteine Proteinase Inhibitors; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005813:Genes, Synthetic; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011480:Protease Inhibitors / Q000235:genetics*; D014158:Transcription, Genetic
D015853:Cysteine Proteinase Inhibitors; D011480:Protease Inhibitors