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. 2021 May 24;9:677867. doi: 10.3389/fcell.2021.677867

FIGURE 1.

FIGURE 1

Production of ZHPV16E6 affibodies. (A) The recombinant plasmids of pET21a(+)/ZHPV16E6 affibodies and pET21a(+)/Zwt affibody were constructed using the NdeI and XhoI restriction sites and further confirmed by DNA sequencing. Amino acid sequences of the ZWT aligned to 3 selected ZHPV16E6 affibodies. The 13 randomized amino acid positions are highlighted in red. Dashes (–) indicate amino acid identities, and black boxes show three α-helical subdomains in the wild-type Z domain. (B) Coomassie Brilliant Blue-stained SDS-PAGE gel of the recombinant proteins in E. coli BL21 induced by 1 mM IPTG. M, protein ladder; 1, empty E. coli BL21(DE3); 2, E. coli BL21(DE3) transformed with pET21a(+) empty vector; 3–6, E. coli BL21(DE3) transformed with pET21a(+)/ZHPV16E61115, pET21a(+)/ZHPV16E61171, pET21a(+)/ZHPV16E6 1235, and pET21a(+)/ZWT plasmid, respectively. The purified ZHPV16E6 affibodies were evaluated by 15% SDS-PAGE (C) and verified by Western blotting (D) M, protein marker; 1, ZHPV16E61115; 2, ZHPV16E61171; 3, ZHPV16E61235; and 4, ZWT.