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. 2020 Nov 12;11:581856. doi: 10.3389/fmicb.2020.581856

FIGURE 1.

FIGURE 1

Expression and analysis of recombinant Nsp9 and Nb6. (A) Size-exclusion chromatography of recombinant Nsp9 on calibrated Superdex 200 increase 10/300 GL column. The target protein was eluted at a retention volume of 13.5 ml, corresponding to approximately 72 kDa as monomer. Characterization of recombinant Nsp9 after two-step purification was analyzed by SDS–PAGE (Inset), M: protein ladder (10–180 kDa, Sorlarbio, Beijing, China), Gel: 12% precast polyacrylamide gel (Item No. NP0341BOX, Thermo Fisher Scientific). (B) Size-exclusion chromatography of recombinant Nb6 on calibrated Superdex 75 10/300 GL column. The target protein was eluted at a retention volume of 13 ml, corresponding to approximately 15 kDa as monomer. Characterization of recombinant Nb6 after two-step purification was analyzed by SDS–PAGE (Inset), M: protein ladder (10–180 kDa, Sorlarbio, Beijing, China), Gel: 12% precast polyacrylamide gel (Item No. NP0341BOX, Thermo Fisher Scientific). (C) Analysis of purified Nb6 binding to Nsp9. Gradient concentrations (62.5 nM–1 μM) of Nb6 in PBS buffer (pH7.4), containing 0.02% tween-20 and 0.1% BSA were passing through the streptavidin biosensor loaded with biotinylated Nsp9, and the signals were recorded.