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. 2022 Jan 19;12:1005. doi: 10.1038/s41598-022-04883-y

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Expression of SARS-CoV-2 VLPs in N. benthamiana. (a) Schematic representation of two vectors for the formation of SARS-CoV-2 VLPs. pBYR2fp-M_IRESE (upper) and pBYR2fp-N_FLAG (bottom) vectors contained a recombinant gene, M-IRES-E (M_IRESE) and N-FLAG (N_FLAG). (b) Schematic representation of the co-expression system process by two vectors using the agroinfiltration method in tobacco plants. (c) Identification of M protein (~ 25 kDa) expression using Western blot analysis in total soluble protein (TSP) extracted from tobacco leaves 3 dpi of pBYR2fp-M_IRESE. Recombinant M protein (15 kDa) derived from E. coli used as a positive control was loaded with 0.25 µg (line 2) and 0.5 µg (line 3), respectively. ME represents samples injected with pBYR2fp-M_IRESE. Western blot analysis under both denaturating (d) and non-denaturating conditions (e) to confirm the co-expression of M, E, and N in TSP. N_FLAG represents samples injected with pBYR2fp-N_FLAG. ME + N_FLAG represents samples co-injected with pBYR2fp-M_IRESE and pBYR2fp- N_FLAG. All images were cropped. See Supplementary Figs. S1–S3 for full size of blot.