DNA origami is used to template the shape and size of capsids generated from purified single subunit capsid proteins. NBThighlight
) has been fused to the C-terminus of the VP1 from the NoV strain Hu/GII.4/Sydney/NSW0514/2012/AU. The particles were quality controlled with dynamic light scattering for particle formation, sodium dodecyl sulfate polyacrylamide gel electrophoresis for protein purity and the residual dsDNA was measured. For the complexation with DNA origami, DNA origami was present in the sample during both disassembly and reassembly of the VLPs.
The SV40 major CP VP1 was disassembled and reassembled by dialysing the assembled VLPs in PBS against 20 mM Tris, 2 mM DTT, 5 mM EDTA and 50 mM NaCl, pH 8.9 for 2 h at 4 °C , after which the EDTA concentration was decreased by an additional dialysis step at 4 °C for 2 h against 20 mM Tris, 2 mM DTT, 2 mM EDTA and 50 mM NaCl, pH 8.9. The concentration was determined based on the absorbance at 280 nm . The DNA origami was transferred into 100 mM HEPES buffer, pH 7.
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