Stringent and reproducible tetracycline-regulated transgene expression by site-specific insertion at chromosomal loci with pre-characterised induction characteristics-1

<p><b>Copyright information:</b></p><p>Taken from "Stringent and reproducible tetracycline-regulated transgene expression by site-specific insertion at chromosomal loci with pre-characterised induction characteristics"</p><p>BMC Molecular Biology 2007;8():30-30.</p><p>Published online 10 May 2007</p><p>PMCID:PMC1884169.</p><p></p>Flow cytometric profiles (y-axis, counts; x-axis, GFP fluorescence [FLH1]) of clones isolated after transfection with plasmids shown in A. For profiles in B (left only), C, D, F and G the green and yellow traces represent cells grown with or without tet, respectively, and the red traces represent GFP-negative cells (Rht14 or HT1080). . Left: Clone 6, the most stringently regulated clone isolated after transfection of pTARG1 into HT2 cells. Right: GFP expression profile of Clone 6 at indicated times after addition of tet. . Four of the most stringently regulated clones isolated after transfection of pTARG2 into HT2 cells. . Three of the most stringently regulated clones isolated after transfection of pTARG3 into HT2 cells. . Effect of AZC on GFP expression in T15, a pTARG3-transfected HT2 clone. Cells were cultured with (green) or without (blue) AZC for 24 h, and then for a further 24 h without AZC, before analysis. . Four of the most stringently regulated clones isolated after transfection of pTARG4 into Rht14 cells. . GFP expression profile of two heterogeneously expressing clones isolated after transfection of Rht14 with pTARG4. . Profile of clone Rht14-10 grown continuously with passaging for 14 d or 42 d. . GFP expression in clone Rht14-19, as measured by immunoblot (top) or flow cytometry (bottom), at the indicated times after addition of tet