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. 2022 Feb 26;134:102186. doi: 10.1016/j.tube.2022.102186

Fig. 1.

Fig. 1

Production, purification, and identification of the recombinant protein rRv0572c. (A) The gene encoding Rv0572c was inserted into pPROEX, to construct prokaryotic expression recombinant plasmid pPROEX-Rv0572c. (B) The recombinant plasmid was confirmed by restriction enzyme digestion with BamH I and Xho I. (C) Expression and purification process of the protein rRv0572c were monitored and confirmed via 15% SDS-PAGE analysis. Lane M, protein marker (kDa); lane 1, recombinant E. coli BL21 (DE3) strain without the induction of IPTG; lane 2, recombinant E. coli BL21 (DE3) strain induced with IPTG for 4 h; lane 3, supernatant of recombinant bacteria induced by IPTG after sonication; lane 4, effluence after binding to the Ni-NTA column; lane 5, effluence after washing with 8 M urea-binding solution; lane 6–9, eluted with elution buffer containing 100 mM imidazole. (D) Identification of purified protein rRv0572c by western blotting with anti-His mouse mAb. Lane M, protein marker (kDa); lane 1, pPROEX empty vector; lane 2, purified rRv0572c probed with specific antibodies.