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Expression of the bovine hypothalamic hormone oxytocin precursor in Escherichia coli
U Nörenberg1, A Brunath, D Richter
1Institut für Zellbiochemie und klinische Neurobiologie, Universitäts-Krankenhaus Eppendorf, Hamburg, F.R.G.
Biochimie
|January 1, 1988
Abstract:
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.