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. 2015 Sep 10;44(1):134–151. doi: 10.1093/nar/gkv889

Figure 1.

Figure 1.

Overexpression, purification and dimerization of Cmr. (A) Cmr was overexpressed in Escherichia coli pMBC370, N-terminal 6x-His tagged Cmr protein expression strain and purified with a HisTrap column. Coomassie brilliant blue stained SDS-PAGE gel showing induction of protein expression. Lanes: M, Molecular weight marker; 1, uninduced pMBC370 lysate; 2, IPTG induced pMBC370 lysate; 3, purified His-Cmr. (B) Western blot using anti-His monoclonal antibody, showing overexpression of Cmr in induced E. coli pMBC370. Lanes: 1, uninduced bacterial cell lysate; 2, induced bacterial lysate. (C) Glutaraldehyde cross-linking of purified Cmr analyzed by western blotting using rabbit anti-Cmr antiserum. Lanes: M, Magic Mark XP Western protein standard; 1, purified Cmr; 2, glutaraldehyde cross-linked Cmr. The molecular weight of Cmr is 26.7 kDa. Note that the overexpressed protein contains a His tag of about 4.4 kDa. Therefore, the recombinant protein migrates at about 31 kDa.