Published June 30, 2019
| Version 1.0
Dataset
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Recombinant PNGaseF production and deglycosylation assays
Description
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.
Notes
The pOPH6:EmPNGaseF vector was a kind gift from Dominic Alonzi, Department of Biochemistry, Oxford University and Sarah Need, Open University. Glycoproteins from Johan Hill, Hubert Brandstätter, Marco Trerotola. PR is funded by a LISCB - UK Wellcome Trust ISSF Grant 204801/Z/16/Z. AL is funded by a PhD Studentship by the Italian Government.
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PR-00006_PNGaseF.pdf
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