Expression of the bovine hypothalamic hormone oxytocin precursor in Escherichia coli.
The bovine oxytocin precursor was expressed in Escherichia coli as a fusion protein by cloning the hormone encoding cDNA in frame behind the replicase gene of the RNA phage MS2. By step-wise extraction with different urea concentrations, the fusion protein was enriched in the 7 M urea fraction and further purified by Sephacryl S-300 chromatography. The oxytocin precursor was cleaved off the fusion protein by cyanogen bromide treatment, chromatographed on FPLC columns and identified by Western blot analysis, using antibodies raised against neurophysin.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D010121:Oxytocin / Q000031:analogs & derivatives* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D014158:Transcription, Genetic
D003062:Codon; D011993:Recombinant Fusion Proteins; D010121:Oxytocin; C046278:oxytocin, Gly-Lys-Arg-