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    Conditional expression systems are of pivotal importance for the dissection of complex biological phenomena. Here, we describe a novel EBV-derived episomally replicating plasmid (p RTS-1) that carries all the elements for conditional expression of a gene of interest via Tet regulation. The vector is characterized by (i) low background activity, (ii) high inducibility in the presence of doxycycline (Dox) and (iii) graded response to increasing concentrations of the inducer. The chicken beta actin promoter and an element of the murine immunoglobin heavy chain intron enhancer drive constitutive expression of a bicistronic expression cassette that encodes the highly Dox-sensitive reverse tetracycline controlled transactivator rt TA2 bi-1 that allows simultaneous expression of two genes, of which one may be used as surrogate marker for the expression of the gene of interest. Tight down regulation is achieved through binding of the silencer t TS One of the most powerful approaches for the dissection of complex biological processes is the ability to individually manipulate the activity of a single gene or a few genes within the context of a cell or a living organism. The tetracycline-regulatable gene expression system (Tet system) has, for a number of reasons, gained wide acceptance. By adapting prokaryotic transcription control elements to the eukaryotic transcriptional machinery, a highly selective regulatory circuit was established ( 1 ), which greatly profited from the exceptional specificity of interaction between operator and repressor of the Tn -encoded tetracycline resistance operon ( 2 ). Graded or threshold response of the tet-controlled gene expression. zithromax label Doxycycline STEMCELL Technologies Tet-Off and Tet-On Gene Expression Systems User Manual - Clontech
     
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