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Revision as of 05:37, 17 October 2018
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Constraction
For the pseudovirus production, we prepared structural gene, and non-structural gene with fluorescence protein gene.
About structural gene, capsid (C), membrane (prM) and envelope (E) are necessary for the formation of viral structure. We prepared pCAG-C and pCAG-prM-E for Serotype I to IV. On the other hand, about non-structural gene with fluorescence protein gene, we prepared EGFP-FMDV2a, DsRed-Express-FMDV2a, ZsYellow-FMDV2a and AmCyan-FMDV2a, and inserted each of them to non-structural genes of dengue virus.
Since these plasmids are introduced into mammalian cells, the promoter and backbone should be optimized for eukaryotic cells. Our team uses pCAG vector which includes CAG promoter to induce high expression of interesting genes in mammalian cells, and pCMV for non-structural region.
Results (Electrophoresis)
As shown in Figure, we successfully obtained each part, ranging from C and prM-E to FP-FMDV2a with non-structural region.
http://parts.igem.org/File:TokyoTech_2018_EPresult.png
Protocol
