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RP - Expression and purification of SUMO E1 (His-Aos1/Uba2 or His-Aos1/Uba2-His)
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Frauke Melchior
ZMBH, Heidelberg, Germany
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The SUMO E1 complex can be purified by two different approaches resulting in E1 with identical activity (Werner et al. 2008). The “classical” protocol describes the purification after coexpression of His-tagged Aos1 and untagged Uba2 in bacteria. A recently developed alternative protocol of our lab involves separate expression and purification of N-terminal His-tagged Aos1 and C-terminal His-tagged Uba2 and subsequent reconstitution of the heterodimeric complex. This procedure has the advantage of higher yields, simplified procedures and the possibility to combine different variants or differently tagged subunits.
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Expression and purification of SUMO E1 (His-Aos1/Uba2 or His-Aos1/Uba2-His)
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