Technical Details:
Expression and Purification of Recombinant and Endgenous 0- GlcNAcase The human O-GlcNAcase cDNA was subcloned into the commercial expression bacterial expression vector PET32a (Novagen, Madison, WI), as was the shorter splice variant (MGEA5 s), to yield fusion proteins with thioredoxi. Both construct were confirmed by sequencing.
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This expression vector is used to express active O-GIcNAcase in E. coli. It produces a fusion protein between thioredoxi and O-GIcNAcase which allows for active O-GIcNAcase to be produced in E. coli.
