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. 2018 Nov 27;19(12):3767. doi: 10.3390/ijms19123767

Figure 1.

Figure 1

Construction of shuttle plasmid and illustration of recombinant baculovirus production and transduction of fish cells. In plasmid pFastBac-ie1-CMV-pf/pFastBac-CMV-ie1-pr, a puromycin–red fluorescent protein (Puro-RFP, pr) or puromycin–green fluorescent protein (Puro-GFP, pf) reporter cassette was driven by a cytomegalovirus (CMV) or white spot syndrome virus (WSSV) immediate-early gene 1 (ie1) (WSSV ie1) promoter, and an adjacent WSSV ie1 or CMV promoter was used to express the gene of interest in the reverse direction. The donor plasmid was transformed to DH10Bac competent cells to generate Bacmid, which was then transfected to Sf-9 cells to obtain the baculovirus. The baculoviral stock was amplified by infecting Sf-9 cells and titrated and transduced into fish cells. MCS, multiple cloning sites. A, poly(A). P1 (P2, P3) viral stock, Passage-1 (-2, -3) viral stock.