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Fig. 1 | Virology Journal

Fig. 1

From: The recombinant pseudorabies virus expressing African swine fever virus CD2v protein is safe and effective in mice

Fig. 1

Construction of recombinant and attenuated strains. a The recombinant fragment was inserted between the 108th and 109th bases at the TK gene's N-terminus. The N-terminal homologous arm was 492 bp, and the C-terminal homologous arm was 466 bp. EGFP was driven by the CMV promoter and Flag-CD2v-Flag by the EF1α promoter. b The attenuated strain of PRV-ΔgE/ΔgI/ΔTK was made by insertion of a single base (T) in the US8 (gE) gene, deletion of base (353 bp) in the US7 (gI) gene, and insertion of two bases (GC) in the UL23 (TK) gene. c The attenuated strain of PRV-ΔgE/ΔgI/ΔTK-(CD2v) was made by insertion of single base (T) in the US8 (gE) gene, deletion of 11 bases (11 bp) in the US7 (gI) gene, insertion of CMV-EGFP-EF1-Flag-CD2v-Flag in the UL23 (TK) gene (which disrupted the TK gene). d Immunofluorescence showed that CD2v was expressed in HEK 293 T cells transfected with pcDNA3.1-EGFP-Flag-CD2v-Flag plasmid or infected with PRV-ΔgE/ΔgI/ΔTK-(CD2v) (red is CD2v labeled with Flag) and white scale represents 50 μm. e Western blotting showed that CD2v bands (89 KD full-length glycosylation band, 45 KD N-terminal glycosylation band, 26 KD C-terminal band, 19 KD N-terminal band) were detected in HEK 293 T cell lysates transfected with pcDNA3.1-EGFP-Flag-CD2v-Flag plasmid or infected with PRV-ΔgE/ΔgI/ΔTK-(CD2v). e Lane 1: protein marker, Lane 2: control, Lane 3: pcDNA3.1 transfected group, Lane 4: pcDNA3.1-EGFP-Flag-CD2v-Flag transfected group, Lane 5: PRV-ΔgE/ΔgI/ΔTK-(CD2v)-infected group

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