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. 2014 Dec;20(12):1963–1976. doi: 10.1261/rna.044354.114

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© 2014 Delvillani et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 1. — Escherichia coli reporter strains of Tet-Trap system. (A) Outline of Tet-Trap system and regulatory elements. Repression by TetR (the sensor) is relieved by TIP2 expression (the signal) and this triggers the transcription of the reporter gene controlled by tetAp. The reporter cassette is integrated between the tonB and yciA loci (nucleotides 1309870–1309872); the tetR cassette between attB and bioB (806594–808521). Details about strain construction are reported in Materials and Methods. Construct elements are represented by an arbitrary scale. Empty boxes, open reading frames; bent arrows, promoters; triangles, intrinsic bidirectional transcription terminators. (B) Reporter gene expression regulation in the Tet-Trap system. The reporter strains carry genes conferring either spectinomycin resistance (aadA:GFP gene; upper part, strain C-5907) or streptomycin sensitivity (rpsL⁺; lower part, strain C-5920) downstream from the tetAp promoter. In C-5920, the endogenous rpsL allele carries the rpsL31 (streptomycin-resistance) recessive mutation. In both strains, the tetR gene is constitutively expressed from P_cat, and transcription from tetAp is repressed unless TIP2 is expressed. (Gray ovals) TetR; (dark gray bar) TIP2 peptide; (thick arrows) mRNAs. Other symbols are as in A.