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. 2012 Apr 23;428(2):136–145. doi: 10.1016/j.virol.2012.04.002

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Copyright © 2012 Elsevier Inc. All rights reserved.

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Fig. 3 — Expression and purification of 3CL-His and confirmation of the cleavage activity of EoV 3CL via anti-3CL polyclonal antibodies. (A) Schematic illustrations of the recombinant fusion proteins 3CL-His, His-Pro_2160–2492, His-Pro_2026–2492, His-Pro_2160–2680-Flag and His-3C3D′. The fusion proteins are depicted as bars. The molecular weights of the cleavage products detected with anti-3CL polyclonal antibodies are indicated. (B) Expression and purification of 3CL-His. Lane M, molecular weight markers; Lane 1, uninduced cell lysate; Lane 2, induced cell lysate; Lane 3, purified 3CL-His. The position of overexpressed 34-kDa 3CL-His is shown. (C) The different fusion proteins 3CL-His (Lane 1), His-Pro_2160–2492 (Lane 2), His-Pro_2026–2492 (Lane 3), and His-Pro_2160–2680-Flag (Lane 4) were subjected to Western blot analysis with anti-3CL polyclonal antibodies. Lane M, molecular weight marker. (D) The fusion proteins 3CL-His (Lane 1) and His-3C3D′ were subjected to Western blot analysis with anti-3CL polyclonal antibodies. Lane M, molecular weight marker.