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Phosphoproteins and phosphopeptides were expressed by E. coli to give yields of 30–200mg of purified protein per litre with an average degree of phosphorylation at multiple sites of 61–83%. The method employed two compatible cohabiting plasmids having low and high copy number, expressing a protein kinase and, more abundantly, the substrate (poly)peptide, respectively. It was used to phosphorylate...
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