Figure 2

Construction of allele replacement vector pKO5.2-C.1-gD-EGFP using LR recombination. (A) Schematic diagram of the pKO5.2-C.1 plasmid vector containing the original KO5 selection markers, the lambda recombination sites (attR) and the negative selection marker for recombination (ccdB gene). (B) Schematic diagram of the plasmid pENTR-gD-EGFP containing attL recombination sites and the EGFP gene in place of the HSV-2 glycoprotein D gene. (C) Schematic diagram of the resulting allele insertion vector following in vitro recombination between pKO5.2-C.1 and pENTR-gD-EGFP. (D) Schematic diagram of the resulting plasmid by-product of the LR recombination reaction.