FIG 2.
Construction of the ANCH2 or ANCHOR3 HCMV BAC used in this study and characterization of viruses derived from the TB40-ANCHOR3 HCMV BAC. Both ANCH2 and the ANCHOR3 HCMV BAC were derived from the TB40-GFP BAC, which contains the GFP gene under the control of a murine CMV immediate-early promoter (pMCMV-MIEP-GFP) inserted in the vector backbone. This gene was replaced by the desired constructs without affecting any other viral gene or sequence. The TB40-ANCH2-Kana HCMV BAC displaying a single ANCH2 target sequence was obtained by exchanging the mCMV-MIEP-GFP gene with an ANCH2-Kanar cassette while the MCMV-MIEP-GFP gene was exchanged by homologous recombination with a Kanar-OR3/GFP-ANCH3 cassette to create the TB40-ANCHOR3 HCMV BAC.