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. 2018 Jun 7;8(13):3544–3558. doi: 10.7150/thno.24607

Figure 1.

Figure 1

Expression and purification of ZHPV16 E7 affitoxin 384 protein. The pET21a (+)/ZHPV16 E7 affitoxin384 plasmid was transformed into E. coli BL21 (DE3). The protein was expressed and purified by Ni-NTA agarose affinity chromatography. (A) Schematic structure of pET21a (+)/ZHPV16 E7 affitoxin384 plasmid. (B) Comassie blue-stained SDS-PAGE gel of the recombinant proteins. M, protein marker; 1, Empty E.coli BL21 (DE3); 2, E.coli BL21 (DE3) transformed with pET21a empty vector; 3, E.coli BL21 (DE3) transformed with pET21a (+)/ZHPV16 E7 affitoxin384 plasmid; 4-5, E.coli BL21 (DE3) transformed with pET21a (+)/ZHPV16 E7 affitoxin384 plasmid and induced by 1 mM IPTG; 6, E.coli BL21 (DE3) transformed with pET21a (+)/Zwt affitoxin; 7-8, E.coli BL21 (DE3) transformed with pET21a (+)/Zwt affitoxin and induced by 1 mM IPTG. (C) Analysis of the purified ZHPV16 E7 affitoxin384 and Zwt affitoxin recombinant proteins by SDS-PAGE. M, protein marker; 1, ZHPV16 E7 affitoxin384; 2, Zwt affitoxin. (D-F) Confirmation of the expression of ZHPV16 E7 affitoxin384 and Zwt affitoxin recombinant proteins by western blot using the primary antibodies against His tag, PE38KDEL and SPA-Z, respectively. 1, Empty E.coli BL21 (DE3); 2, E.coli BL21 (DE3) transformed with pET21a empty vector; 3, E.coli BL21 (DE3) transformed with pET21a (+)/ZHPV16 E7 affitoxin384 plasmid and induced by 1 mM IPTG; 4, E.coli BL21 (DE3) transformed with pET21a (+)/Zwt affitoxin and induced by 1 mM IPTG.