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. 2018 Feb 8;9:181. doi: 10.3389/fimmu.2018.00181

Figure 1.

Figure 1

The generation and verification of virus-like particle (VLP)-conjugated CspZ proteins. (A) Purified wild-type CspZ or CspZ-Y207A/Y211A with an exposed a sulfhydryl group and virus-like particles (“VLP”) generated from Qβ phage were mixed with Succinimidyl 6-((beta-maleimidopropionamido) hexanoate) (“SMPH”) to crosslink VLP and each of these CspZ proteins (“VLP-CspZ”). (B) The sizes and purity of each conjugate were determined by a 15% SDS-PAGE. The molecular marker (kD) is in lane 1, followed by preparations of VLP (lane 2, “VLP”), CspZ (lane 3, “CspZ”), VLP-CspZ (lane 4, “VLP-CspZ-WT”), and VLP-CspZ-Y207A/Y211A (lane 5, “VLP-CspZ-Y207/Y211A”). The arrows identify known protein products as indicated. (C) Representative images of VLPs generated in this study. EM analysis of freshly purified VLP (“VLP”), VLP-CspZ (“VLP-CspZ-WT”), or VLP-CspZ-Y207A/Y211A (“VLP-CspZ-Y207A/Y211A”) following negative staining with 1% uranyl acetate. Scale bar = 100 nm.