Use of Recombination-Mediated Genetic Engineering for Construction of Rescue Human Cytomegalovirus Bacterial Artificial Chromosome Clones

Figure 3

Toledo ROI Rescue BAC Clone Construction. Since the ROI rescue plasmid contains restriction enzyme sites (a), the plasmid is digested by restriction endonucleases (b), leading to isolation of the rescue cassette (c). Electrocompetent and recombination-activated DY380 harboring the ROI Δ mutant Toledo BAC (d) are used for electroporation with the rescue cassette (e). Upon homologous recombination, the ROI is inserted into the ROI Δ mutant Toledo BAC, generating the ROI-Rescued Toledo BAC (f).