A gene expression system offering multiple levels of regulation: the Dual Drug Control (DDC) system-3

<p><b>Copyright information:</b></p><p>Taken from "A gene expression system offering multiple levels of regulation: the Dual Drug Control (DDC) system"</p><p>BMC Biotechnology 2004;4():9-9.</p><p>Published online 29 Apr 2004</p><p>PMCID:PMC420247.</p><p>Copyright © 2004 Sudomoina et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p>imerizer only; scale is on the right Y-axis) and the SEAP gene (tTA-dependent transcription expression responsive to dimerizer AND tetracycline; scale is on the left Y-axis). All cells contain pCN-RS/ZF3, pZI-hGH-2, pZBS-Tet-OFF, and pBI-EGFP-SEAP. See Results for details of manipulations A-D; briefly, A was grown in continuous dimerizer-/tetracycline+; B-D were initially in dimerizer+/tetracycline+. At 48 hours after transfection with plasmids, A and B were harvested, while C and D were grown for 48 more hours in dimerizer+/tetracycline+ (C) or dimerizer+/tetracycline- (D) medium before harvesting. Results of one typical experiment, with four parallel transfections for each data point, are shown. Each experiment was repeated at least three times

A gene expression system offering multiple levels of regulation: the Dual Drug Control (DDC) system-1

<p><b>Copyright information:</b></p><p>Taken from "A gene expression system offering multiple levels of regulation: the Dual Drug Control (DDC) system"</p><p>BMC Biotechnology 2004;4():9-9.</p><p>Published online 29 Apr 2004</p><p>PMCID:PMC420247.</p><p>Copyright © 2004 Sudomoina et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</p>; the total amount of plasmid DNA was constant at 0.5 μg per test point. As graphically represented, at each apex of the triangle shown, only one of the test plasmids is present (100% of mix), while opposite the matrix this plasmid is absent (0% of mix), with the two other plasmids present in equal proportions. White circles indicate ratios of plasmids evaluated for degree of induction of SEAP activity. Degree of induction reflects difference between growth without dimerizer and with tetracycline (OFF), versus with dimerizer and without tetracycline (ON). Based on the ON:OFF ratios obtained, the software program "Statistica" modeled the optimal response surface, shown as a contour plot on the triangle. Faded edges of the triangle represent limitations of the model, because it is meaningless whenever one of the components is absent. The optimal ratio for plasmid transfection with the DDC system was 1.5:1.5:1 (pCN-RS/ZF3:pZBS-Tet-OFF:pBI-EGFP-SEAP). This contrasted with a ratio of 3:1 (activator proteins:reporter) used for the original ARIAD system