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RP - Expression and purification of Ubc9 (SUMO E2)
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Sumoylation is a process which requires an enzymatic cascade involving an E1 activating enzyme, an E2 conjugating enzyme and in most cases in vivo an E3 ligase enzyme. Reconstitution of the sumoylation reaction in vitro has the obvious advantage that SUMO isopeptidases, E3 ligases and competing substrates present in eukaryotic extracts can be avoided.
Here, we provide a complete protocol describing the expression and purification of the recombinant E2 enzyme of the SUMO pathway, called Ubc9. Notice that this protocol allows the purification of a non-tagged Ubc9, avoiding any consequences due to the presence of a tag.
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Expression and purification of Ubc9 (SUMO E2)
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