Secretion of human interferon-alpha induced by using secretion vectors containing a promoter and signal sequence of alkaline phosphatase gene of Escherichia coli.
We constructed a new vector containing the promoter and the signal sequence of E. coli phoA gene, the structural gene for the periplasmic alkaline phosphatase. One of the most useful characteristics of this vector is the unique HindIII restriction site located just at the end of the phoA signal sequence. This restriction site was generated by oligonucleotide-directed site-specific mutagenesis without changing the amino acid sequence of the signal peptide. Any kind of foreign structural gene can be easily inserted into the HindIII site by using synthetic oligonucleotides to construct a hybrid gene which has neither an extra sequence nor a deletion between the phoA signal sequence and the foreign structural gene. Human alpha-interferon gene was inserted into this HindIII site. When this hybrid gene was expressed under the control of the phoA promoter region, a low but significant activity was recovered in the cold water wash of the cells after an osmotic shock procedure.
D000469:Alkaline Phosphatase / Q000235:genetics; D004274:DNA, Recombinant; D004926:Escherichia coli; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D007370:Interferon Type I / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D001616:beta-Galactosidase / Q000235:genetics
D004274:DNA, Recombinant; D007370:Interferon Type I; D010455:Peptides; D021382:Protein Sorting Signals; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase