Recombinant PNGaseF production and deglycosylation assays
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Recombinant PNGaseF production and deglycosylation assays. Open Lab notebook. 1.5 L of BL21 E.coli cells harbouring the pOPH6:EmPNGaseF vector encoding a His-tagged construct of Elizabethkingia miricola PNGase F (EmPNGaseF, Uniprot P21163, PNGF_ELIMR) were used to express the protein. EmPNGAse F was purified from the E.coli lysate by immobilised metal affinity and size exclusion chromatography and its activity assayed against 5 distinct glycoproteins.
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2019-06-30
