Expression of the bovine hypothalamic hormone oxytocin precursor in Escherichia coli |
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| Authors: | U N?renberg A Brunath D Richter |
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| Institution: | Institut für Zellbiochemie und klinische Neurobiologie, Universit?ts-Krankenhaus Eppendorf, Hamburg, F.R.G. |
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| 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. |
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