Cloning a synthetic gene for human stefin B and its expression in E. coli.
A gene coding for human stefin B was synthesized by the solid-phase phosphite method and cloned in the pUC8 cloning vector. The insert with the verified DNA sequence was subcloned into two expression vectors and expressed in E. coli as a fusion protein with beta-galactosidase and as a native protein. The CNBr cleaved fusion protein and the native recombinant stefin B were inhibitory to papain and reacted with antibodies against human stefin B.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D055313:Cystatin B; D015891:Cystatins; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005813:Genes, Synthetic; D006801:Humans; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000235:genetics*; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification; D014158:Transcription, Genetic
C524994:CSTB protein, human; D015891:Cystatins; D011480:Protease Inhibitors; D011506:Proteins; D011994:Recombinant Proteins; D055313:Cystatin B