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. 2020 Mar 4;10:4017. doi: 10.1038/s41598-020-61103-1

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Generation and characterization of bicistronic lentiviral vectors expressing flaviviral structural proteins. (A) JEV CprME was cloned into a lentiviral vector that included an IRES sequence followed by the Zika NS2B-3 protease. 293 T cells were transfected with the JEV CprME construct alone or the bicistronic JEV lentiviral construct. Culture supernatants were harvested and VLPs analyzed for JEV E protein and Capsid protein secretion via western blotting. (B) YFV CprME was cloned into a lentiviral vector as above and VLP secretion determined in the culture supernatants by western blotting for E and Capsid protein. (C) CHIKV C-E3-E2-E1 genes were cloned into a lentiviral vector and VLP secretion determined in the culture supernatants by western blotting for E1-E2 protein. Images were analyzed using GENETOOLS gel analysis Software version 4.03 (f). (Syngene, https://www.syngene.com/software/genetools-automatic-image-analysis/).