Research Paper Volume 17, Issue 1 pp 161—169

Figure 1. Diagrammatic representation of main intermediate constructs generated during the construction of HD-RAd-STEMCCA-GFP-Tet-Off. (A) Shows the base plasmid, pLPBL-BD-tTA-GFP, from which we departed in the generation of the above adenovector. It harbors the CMV1-TRE-CMV2 regulatable promoter flanked on one side by the gene for GFP and by a multiple cloning site (MCS) on the other side. Separately, the plasmid also harbors the gene for the chimeric tetracycline transactivator (tTA) protein, created by fusing one protein, TetR (tetracycline repressor), with the activation domain VP16, from the Herpes Simplex Virus. This construct is under the control of the human cytomegalovirus (CMV) promoter. (B) Displays the above plasmid now harboring the STEMCCA tandem, which was cloned into the MSC site. (C) Shows the full STEMCCA construct (including all associated regulatory components) cloned into the HD genomic plasmid which includes a bacterial sequence between its ITRs, flanked by PmeI sites. (D) Illustrates the basic components of HD-RAd-STEMCCA-GFP-Tet-Off genome after removing the bacterial sequence from pC4HSU-tTA-STEMCCA-GFP. For further details see Figure 2A legend and Ref. 15.4.