Chemical synthesis of a human interferon-alpha 2 gene and its expression in Escherichia coli.
A 511-base pair DNA fragment encoding human interferon-alpha 2 has been chemically synthesised and expressed from a lac UV5 and a synthetic trp promoter in Escherichia coli. The synthesis involved preparation of 68 oligodeoxyribonucleotides and their enzymic ligation. The product expressed from the trp promoter system had high antiviral activity and displayed biological effects similar to those of Namalwa interferon on natural killer cell activity and in a Daudi cell growth inhibition assay. E.coli minicells containing plasmid DNA with the synthetic IFN-alpha 2 gene under trp promoter control produce a protein with the same electrophoretic mobility as a sample of authentic IFN-alpha 2. The protein from E.coli cross-reacts with the monoclonal antibody NK-2 and was readily purified, close to homogeneity, by immunoadsorption chromatography on NK-2 sepharose.
D000595:Amino Acid Sequence; D001483:Base Sequence; D001681:Biological Assay; D002460:Cell Line; D003001:Cloning, Molecular; D004261:DNA Replication / Q000187:drug effects; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D007370:Interferon Type I / Q000138:chemical synthesis / Q000235:genetics* / Q000494:pharmacology; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000138:chemical synthesis; D010957:Plasmids
D007370:Interferon Type I; D009841:Oligonucleotides; D004262:DNA Restriction Enzymes