pBI Tet

The pBI Tet Vector is a response plasmid that can be used to express two genes of interest from one bidirectional tet-responsive promoter (Pbi-1; 1) in Clontech’s Tet-On® and Tet-Off® Gene Expression Systems and Cell Lines (2). The Tet Expression Systems and Cell Lines give researchers ready access to the tetracycline-regulated expression systems described by Gossen & Bujard (3; Tet-Off) and Gossen et al. (4; Tet-On). The pBI Tet Vector contains the bidirectional promoter Pbi-1 which is responsive to the tTA and rtTA regulatory proteins in the Tet-Off and Tet-On systems, respectively. Pbi-1 contains the Tet-responsive element (TRE), which consists of seven copies of the 42-bp tet operator sequence (tetO). The TRE element is between two minimal CMV promoters (PminCMV), which lack the enhancer that is part of the complete CMV promoter. Consequently, Pbi-1 is silent in the absence of binding of TetR or rTetR to the tetO sequences. PminCMV-1 and PminCMV-2 control the expression of two separate genes of interest. Note that the cloned inserts must have an initiation codon. In some cases, addition of a Kozak consensus ribosome binding site (5) may improve expression levels; however, many cDNAs have been efficiently expressed in Tet systems without the addition of a Kozak sequence.

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