Chemical synthesis of a gene for human stefin A and its expression in E. coli.
A DNA containing the coding sequence for the human cysteine proteinase inhibitor stefin A was obtained by enzymic ligation of chemically synthesized deoxyoligonucleotides, using the Khorana ligation method. The 306-bp synthetic gene carries signals for the initiation and termination of its translation. The gene was expressed in E. coli using a cytoplasmic expression vector and stefin A was secreted under the control of the E. coli alkaline phosphatase signal sequence, respectively. The secreted hybrid protein was shown to exhibit biological properties similar to the native protein isolated from human plasma.
Strauss
M
M
Institut für Biochemie, Technische Hochschule Darmstadt.
Stollwerk
J
J
Lenarcic
B
B
Turk
V
V
Jany
K D
KD
Gassen
H G
HG
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D055313:Cystatin B; D015891:Cystatins; D004247:DNA / Q000138:chemical synthesis*; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005813:Genes, Synthetic; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
C524994:CSTB protein, human; D015891:Cystatins; D009838:Oligodeoxyribonucleotides; D011506:Proteins; D011994:Recombinant Proteins; D055313:Cystatin B; D004247:DNA