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Review
. 1999 Feb 2;96(3):797-9.
doi: 10.1073/pnas.96.3.797.

Tet B or not tet B: advances in tetracycline-inducible gene expression

H M Blau  1 , F M Rossi
Affiliations
Review

Tet B or not tet B: advances in tetracycline-inducible gene expression

H M Blau et al. Proc Natl Acad Sci U S A. .
No abstract available

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Figures

Figure 1
Figure 1
(A) Graded transcriptional response to tet at the single cell level. A dose-dependent response to increasing concentrations of an inducer usually can be observed by measuring expression of a reporter gene in bulk assays. Such assays, however, cannot distinguish between two alternative mechanisms operating at the single cell level. In case 1, each cell could respond to intermediate concentrations of inducer by expressing an intermediate amount of gene product (graded response), or in case 2, an intermediate concentration of inducer could result in a fixed maximal level of gene product in a subset of the cells, with the number of cells expressing the gene depending on the concentration of inducer in the medium (threshold response). Case 1 applies to the tet system (3). (B) The need for tetR dimerization specificity. When attempting to coexpress tetR fusion proteins containing different functional domains such as repressor domains (symbolized in the top row by the “do not enter” sign), and activator domains (symbolized by the “go” sign); or DNA binding domains with distinct specificity (symbolized in the middle row by the blue and red “feet”), both productive homodimers and nonproductive heterodimers will form. Nonproductive interactions can be avoided by engineering specific dimerization domains in the tetR portion of the fusion proteins (symbolized by the black and green color midsections in the bottom row). (C) Increasing the dynamic range of the tet system. The basal level of expression of genes under tet control can be reduced without affecting the fully induced level by coexpressing in the same cells a repressor and an activator that respond oppositely to dox and that cannot heterodimerize because of different dimerization domains. The black DNA binding domain symbolizes the wild-type tetR (that binds the tetO in the absence of dox), whereas the white DNA binding domain symbolizes the “reverse” tetR (that binds the tetO in the presence of dox). (D) Independent expression of two different genes. By coexpressing two tetR-based activators that contain DNA binding domains with distinct specificity, respond oppositely to dox, and do not heterodimerize, two independent genes can be regulated by the same inducer. Because of the characteristic dose response of the wild-type and “reverse” tetR, the expression of each gene can be turned off at an intermediate concentration of dox and activated at markedly different dox concentrations.
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References

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