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. 2013 May 20;12:49. doi: 10.1186/1475-2859-12-49

Figure 2.

Figure 2

Yellow fluorescent protein variants and Sec-pathway export. A) Schematic representation of yellow fluorescent protein variants targeted to the periplasm via the Sec-pathway. Standard eYFP targeted to the periplasm does not result in significant levels of protein. In contrast engineered secretion optimized sfYFP yields active fluorescent protein in the bacterial periplasm (cartoon of eYFP generated from pdb 3V3D with mutated residues as grey stick representation. B) SDS-PAGE of Strep-tactin affinity purified eYFP (pTDpelB-C_eYFPTwinStrep) (lane 1) and sfYFP (pTDpelB-C_sfYFPTwinStrep) extracted from the periplasm of E.coli BL21 (lane 2) and eYFP (pTDpelB-C_eYFPTwinStrep) (lane 3) and sfYFP (pTDpelB-C_sfYFPTwinStrep) extracted from whole cells (lane 4). M = PageRuler Unstained Broad Range Protein Ladder (ThermoScientific) with 70, 50, 40, 30, 20 kDa bands.