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Figure 5 | BMC Biology

Figure 5

From: Eukaryotic protein production in designed storage organelles

Figure 5

PBs isolation by density and Zera-fusion proteins recovery. (A-C) Protein analysis of fractions collected from a sucrose gradient of Zera-EGF tobacco leaf homogenates. (A) Immunoblot (αR8 antibody) shows Zera-EGF primarily in the dense F3 fraction (1.18 to 1.26 g/cm3 interface, arrow). (B) Immunoblot using anti-BiP antibody indicates that the soluble ER chaperone co-sediments with Zera-EGF in the F3 fraction. (C) Silver stained SDS-PAGE of sucrose fractions showing the enrichment of Zera-EGF in the F3 fraction. (D) Subcellular fractionation of homogenates of T. reesei expressing Zera-GFP (right) and GFP-KDEL (left). The analysis of fractions by Coomassie staining shows highly concentrated Zera-GFP in the dense fraction F3 (1.18 to 1.26 g/cm3 interface, arrow). (E) Protein analysis by gel electrophoresis and silver stain of fractions collected from sucrose gradients of Zera-EGF (left) and EGF (right) Sf9 expressing cells. Recombinant Zera-EGF fusion is concentrated in the dense fraction F3 (arrow) where PBs sediment.

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