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. 2015 Apr 23;11(6):1456–1464. doi: 10.1080/21645515.2015.1037057

Figure 1.

Figure 1.

Construction, purification and identification of the recombinant fusion protein CTT3H. The gene encoding the polyprotein CTT3H composed of 5 antigens CFP10, TB10.4, TB8.4, Rv3615c, and HBHA of M. tuberculosis was cloned into pET30b, resulting in the recombinant plasmid pET30b-CTT3H (A). The constructed recombinant plasmid pET30b-CTT3H was verified by enzyme digestion with NdeI and XhoI (B). pET30b-CTT3H transformed E. coli BL21(DE3) strain was induced with IPTG. The expression and purification process of polyprotein CTT3H was monitored and confirmed by 12% SDS-PAGE, respectively (C). Lane M, Pre-stained protein ladder of Fermentas; Lane 1, pre-induced; Lane 2, post-induced; Lane 3 and 4, flow-through fraction of His-bind resin; Lane 5 and 6, fraction eluted with 500 mM imidazole, Lane 7, the final products. The purified polyprotein CTT3H was also confirmed by western blotting with an anti-His 6 mouse monoclonal antibody or a mouse polyclonal serum against CTT3H, respectively (D).