FIG. 1.
Cloning strategy for the production of the K181Perth-based BAC, pARK25. The BAC cassette contained within the pAJR02 plasmid was inserted into the K181 strain of MCMV in tissue culture by homologous recombination to produce the BAC pARK14 with a deletion in the genes m07 to m12 (nt 6522 to 12488). Repair of pARK14 was achieved by two-step allele replacement using the pAJR10 plasmid to produce the full-length BAC, pARK25. The EcoRI (E) and HindIII (H) RE sites, as well as the gpt and EGFP genes of the BAC cassette, are shown. Repair of pARK14 to pARK25 produces a 249-bp repeat region (black boxes) flanking the BAC cassette that allows for homologous recombination and reconstitution of wt genotype (vARK25) upon virus rescue. Fragment sizes (in kilobases) are shown in boldface type above or below the fragment lines.