Fig. 6: Characterisation of the expression of heterologous proteins (AeBlue and HPV16 capsid L1) via multi-copy genome integration (MI) using P_BTS1-RPL25-driven in vivo gene amplification.

a Schematic of genetic vectors used to express AeBlue and HPV16 L1. b cells harbouring an empty 2μ, the amplifiable AeBlue construct (MI), AeBlue-and-HPV16-L1 2μ plasmid, and amplifiable AeBlue-and-HPV16-L1 construct (MI). Cells were grown in MES-buffered YNB medium with 20 g l⁻¹ glucose and collected at 72 h, or were grown in YP medium with 20 g l⁻¹ galactose to OD₆₀₀ = ∼20. c Ultracentrifugation of the supernatant on an iodixanol gradient to separate a band containing HPV16-L1 virus-like particles (shown by orange arrow), and transmission electron microscopy confirming the presence of HPV16-L1 virus-like particles (VLPs). d SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) for whole-cell lysates, lysate supernatant, and lysate pellets of yeast samples in b, and VLPs sample from c. Experimental repetition is not done for c and d. Numbers in b–d are for sample cross-reference. The bands d1, d2, d3, and d4 are analysed using a LC-MS/MS-based proteomic method (Supplementary Method 1), and the data are available in Supplementary Data 1 (d1), Supplementary Data 2 (d2), Supplementary Data 3 (d3), and Supplementary Data 4 (d4). Source data for VLPs in d are provided as a Source Data File.