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. 2021 Sep 2;11:17542. doi: 10.1038/s41598-021-97038-4

Figure 1.

Figure 1

FLAG tag insertion at three positions of DENV E protein domain II in DENV-1 VLP expression plasmid. (a) Amino acid positions of FLAG tag insertion in E protein of DENV-1 are shown. (b) The 3D structure of DENV-1 E protein (aa 2–404 region) was constructed and three positions of FLAG tag insertion were shown. Circle symbols indicate FLAG tag insertion positions at aa 77/78, 223/224 and 273/274. Three domains in E protein were distinguished in red (domain I), yellow (domain II) and blue (domain III). (c) Expression of DENV-1 E protein with FLAG tag insertion from prME plasmids FL-223, -273 and -77 in HEK293T cells (left) and their culture supernatants (right) were compared by western blotting (WB). Monoclonal antibodies against FLAG tag (FLAG), flavi group antigen (E protein), and GAPDH were used as 1st antibodies. Non-transfected HEK293T cell lysate and culture supernatant samples were added as a negative control. Because of difference in expression efficiency among prME plasmids tested, amounts of samples applied for WB which produced from FL-273 were one-hundredth (cell lysate) and one-tenth (supernatant) compared to those from FL-223 and FL-77. The amounts of cell lysates applied were 3 mg for FL-223, FL-77 and HEK293T and 0.03 mg for FL-273. Uncropped images of western blotting pictures are included in supplementary Fig. 1.

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