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Fig. 4 | New Zealand Journal of Forestry Science

Fig. 4

From: The occurrence of progesterone 5β-reductase is not limited to the angiosperms: a functional gene was identified in Picea sitchensis and expressed in Escherichia coli

Fig. 4

Purification steps to isolate recombinant P5βR protein ABK24388.1 of P. sitchensis from E. coli host cells using an N-terminal 6× His tag SUMO protein fusion. Five micrograms of total protein per lane was analysed by a 12 % SDS-PAGE. Lane 1: Standard proteins of indicated molecular weights (Thermo #26612, Thermo Scientific, Darmstadt, Germany). Lane 2: Cell-free protein extract of E. coli BL21DE3/pLysS/ pET28 M-SUMO3-ABK24388.1 prepared 8 h after the addition of IPTG. Lane 3: Protein fraction eluted from the first Ni-NTA affinity column. Lane 4: Protein fraction eluted from the first Ni-NTA affinity column following incubation with SenP2 protease. Lane 5: Flow-through of the second Ni-NTA affinity column. Lane 6: Collected major peak fractions obtained after anion-exchange chromatography on a Resource Q column

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