pTACT1
(Plasmid #24779)

• Depositing Lab: Tanya Parish
• Publication: Carroll et al Appl Environ Microbiol. 2005 Jun . 71(6):3077-84.

Backbone

• Vector backbone: pMV206
• Backbone size w/o insert (bp): 4120
• Vector type: Mycobacterial shuttle vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: TetR
• Species: Bacterial
• Insert Size (bp): 900
• Entrez Gene: tetR (a.k.a. pEK516_p06)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: PstI (not destroyed)
• 5′ sequencing primer: n/a

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Ambrose Cheung Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

There are a few mismatches between depositor's sequence and the quality control sequence Addgene has obtained. The region of mismatch and gaps is derived from the parental vector and not the important part of the vector.

Infect Immun. 2001 Dec;69(12):7851-7.
Evaluation of a tetracycline-inducible promoter in Staphylococcus aureus in vitro and in vivo and its application in demonstrating the role of sigB in microcolony formation.

Bateman BT, Donegan NP, Jarry TM, Palma M, Cheung AL.

Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Abstract

An inducible promoter system provides a powerful tool for studying the genetic basis for virulence. A variety of inducible systems have been used in other organisms, including pXyl-xylR-inducible promoter, the pSpac-lacI system, and the arabinose-inducible P(BAD) promoter, but each of these systems has limitations in its application to Staphylococcus aureus. In this study, we demonstrated the efficacy of a tetracycline-inducible promoter system in inducing gene expression in S. aureus in vitro and inside epithelial cells as well as in an animal model of infection. Using the xyl/tetO promoter::gfp(uvr) fusion carried on a shuttle plasmid, we demonstrated that dose-dependent tetracycline induction, as measured by bacterial fluorescence, occurred in each of the above environments while basal activation under noninduced conditions remained low. To ascertain how the system can be used to elucidate the genetic basis of a pathogenic phenotype, we cloned the sigB gene downstream of the inducible promoter. Induction of SigB expression led to dose-dependent attachment of the tested strain to polystyrene microtiter wells. Additionally, bacterial microcolony formation, an event preceding mature biofilm formation, also increased with tetracycline induction of SigB.

PMID: 11705967 [PubMed - indexed for MEDLINE]PMCID: PMC98881Free PMC Articl