Journal: bioRxiv
Article Title: Generation of broad-spectrum vaccine platforms against tick-borne flaviviruses using ancestral sequence reconstruction
doi: 10.1101/2025.10.31.685878
Figure Lengend Snippet: (A) Plasmid constructs were designed to function as vaccine platforms for high level expression of ancestral TBF envelope antigen using an alphavirus infectious clone platform (SINV WT). A gBlock consisting of the TBF +E sequence was inserted into a Sindbis virus infectious clone following a 3’ subgenomic promoter (3’ SGP) via restriction enzyme digestion and Gibson Assembly (SINV +E). (B) Viral titers of passage 1 (p1) from plasmid-transfected BHK-21 cells. BHK cells were infected at MOI=0.01. Supernatant was collected at indicated times and viral titers were determined by standard plaque assay. (C) Cycle threshold (Ct) values of p1 SINV WT (red) and SINV +E (blue) stocks probing for the ancestral TBF E (ASR TBF E).
Article Snippet: Briefly, SINV WT plasmid was digested with XbaI restriction enzyme (RE) (New England Biolabs, R0145S) to generate a linear product and gel extracted.
Techniques: Plasmid Preparation, Construct, Expressing, Sequencing, Virus, Transfection, Infection, Plaque Assay