Expression of the chemically synthesized coding region for the cytotoxin alpha-sarcin in Escherichia coli using a secretion cloning vector.
The coding region for the cytotoxin alpha-sarcin from Aspergillus giganteus has been chemically synthesized by the ligation of 19 overlapping oligodeoxyribonucleotides. An Escherichia coli clone producing the cytotoxin was constructed by inserting the synthesized gene directly downstream to the region coding for the signal peptide of the OmpA protein (a major outer membrane protein of E. coli), using the secretion cloning vector pIN-III-OmpA2. The enzyme encoded by the chemically synthesized gene expressed in E. coli displayed properties identical to those of native alpha-sarcin isolated from A. giganteus with respect to its chemistry, antigenicity and ribonucleolytic activity in qualitative assays.
Institut für Biochemie, Freie Universität Berlin.
P P
PP
Henze
U
U
Hahn
V A
VA
Erdmann
N
N
Ulbrich
D000595:Amino Acid Sequence; D001483:Base Sequence; D003854:Deoxyribonucleotides / Q000138:chemical synthesis; D004722:Endoribonucleases; D004926:Escherichia coli / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015870:Gene Expression; D005813:Genes, Synthetic / Q000502:physiology*; D005822:Genetic Vectors / Q000502:physiology*; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000138:chemical synthesis; D010957:Plasmids / Q000502:physiology; D011500:Protein Synthesis Inhibitors / Q000302:isolation & purification / Q000378:metabolism
D003854:Deoxyribonucleotides; D005656:Fungal Proteins; D009841:Oligonucleotides; D011500:Protein Synthesis Inhibitors; C014801:alpha-sarcin; D004722:Endoribonucleases