Stringent and reproducible tetracycline-regulated transgene expression by site-specific insertion at chromosomal loci with pre-characterised induction characteristics-5
<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>-mediated insertion of pIN2-neoSCE or pIN2-neoR52. . Ethidium bromide stained agarose gel of electrophoresed PCR products generated with the primers O2 and O5 on cell pellets of three pIN2-neoSCE- and three pIN2-neoR52-transfected Rht14-10 derivatives. Negative (-) and positive (+) controls were, respectively, no cells and cells (TetNeo, see methods) with an integrated tet-regulated neo cassette (expected product: 944 bp). Clones chosen for flp-mediated deletion (10IN-SCE.1 and 10IN-R52.1) are arrowed. . As in B but after Flp mediated deletion. . Ethidium bromide stained agarose gel of electrophoresed PCR products generated with the primers O2 and O6 on clones derived from 10IN-SCE.1 (1–12, left) and primers O2 and O7 on clones derived from 10IN-R52.1 (1–12, right). Control reaction contained no cells (-) or genomic DNA generated from pools of flp-treated pIN2-neoSCE/pIN2-neoR52 cells (+). Clones chosen for further analysis (10IN-SCE.1flp1 and 10IN-R52.1flp1) are arrowed. . Immunoblot analysis of I-I and Rad52 expression in Rht14-10 and, respectively in 10IN-SCE.1flp1 or 10IN-R52.1flp1 at the indicated times after removal or tetracycline
