A resolvase-based screen to identify in vivo repressed genes. (A) Schematic illustration of the genetic components of the TetR-controlled in vivo expression technology (TRIVET). Chromosomal sequences are highlighted in light gray; tetR-phoA-cat (tpc) cassette in black; integrated pTRIVET in dark gray; and the res cassette parts in open shapes. The tpc cassette is integrated into a V. cholerae hypothetical geneX via Tn10 mutagenesis; resulting in a transcriptional fusion of tetR and phoA to the chromosomal promoter of geneX. Subsequently; pTRIVET is integrated downstream of lacZ. Thus; the expression level of TetR via the geneX promoter dictates the expression of TnpR; which catalyzes the excision of the res cassette. The IS10 elements and constitutive cat promoter of the tpc cassette; the TetR-controlled tnpR; mobilization (mob); origin of replication (oriR6K); and ApR (bla) regions of pTRIVET; as well as the gene for KmR (neo); SucS (sacB); and the target sites of resolvase (res) of the res cassette are indicated. (B) Shown are the effects of iron availability on resolution frequency (red) and alkaline phosphatase (PhoA) activity (black) of strain Vc_res1_TRIVET irgA

Cat. #160861

TetR-controlled recombination-based in vivo expression technology (TRIVET) vector

Cat. #: 160861

Sub-type: Reporter

Availability: Please enquire for quantities and pricing

Target: Cholera

Bacterial Resistance: pTRIVET - ampicillin, tpc reporter cassette - chloramphenicol

Selectable Markers: PhoA activity

This fee is applicable only for non-profit organisations. If you are a for-profit organisation or a researcher working on commercially-sponsored academic research, you will need to contact our licensing team for a commercial use license.

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Contributor

Inventor: Stefan Schild

Institute: University of Graz

Tool Details

Target Details

Application Details

References

Tool Details

*FOR RESEARCH USE ONLY (for other uses, please contact the licensing team)

Tool name: TetR-controlled recombination-based in vivo expression technology (TRIVET) vector
Alternate name: TRIVET
Research fields: Microbiology
Tool sub type: Reporter
Bacterial resistance: pTRIVET - ampicillin, tpc reporter cassette - chloramphenicol
Selectable markers: PhoA activity
Description: This system is based upon recombination-based in vivo expression technology (RIVET) and allows the detection of transient gene silencing events even in a subset of a complex, heterogenous bacterial population. It requires all three components; the tpc reporter cassette, pTRIVET and pTRIVET1. It can be used as a random approach to identifyÄËĂÂÄšĹĽin vivoÄËĂÂÄšĹĽrepressed genes during intestinal colonization and as a specific approach to study conditional transcriptional silencing of a gene of interest. The protocol for which can be found at PMID: 33659431.
Additional notes: Vibrio cholerae gene expression

Target Details

Target: Cholera

Application Details

Application notes: This system is based upon recombination-based in vivo expression technology (RIVET) and allows the detection of transient gene silencing events even in a subset of a complex, heterogenous bacterial population. It requires all three components; the tpc reporter cassette, pTRIVET and pTRIVET1. It can be used as a random approach to identifyâ€¯in vivoâ€¯repressed genes during intestinal colonization and as a specific approach to study conditional transcriptional silencing of a gene of interest. The protocol for which can be found at PMID: 33659431.

References

33659431
Cakar et al. 2018. Proc Natl Acad Sci U S A. 115(10):E2376-E2385. PMID: 29463743.