Available technologies

Improved Mycobacterial Tetracycline Inducible Vectors

Reference number: 7722, 7726, 7727, 7728

A range of improved tetracycline inducible vectors to study the function of essential mycobacterial genes.

Proposed use

Conditional gene expression systems are powerful tools for studying essential genes in bacteria. Tetracycline and its analogues are ideal inducer molecules as they regulate gene expression at very low concentrations (nM range), show good bio-availability (ie. they can penetrate both mycobacterial and animal cells), are non-toxic at the necessary levels and are stable over the required length of study.

Problem addressed

There is a lack of genetic tools available to researchers to study the function of essential mycobacterial genes, we have therefore constructed a range of improved tetracycline inducible vectors. We have shown that changing the terminators upstream of the inducible promoter does not affect background transcription, and that repressor binding to the operator is a more critical step in controlling regulation. We have eliminated further read-through problems by changing the backbone vector used. The TetRO promoter can be induced in a dose-dependent manner by tetracycline, anhydrotetracycline and doxycycline.

Technology overview

Plasmid Description
pMEND-Lx pMIND-Lx with Gly40 to Thr40 change in TetR
protein; episomal
pMEND-Lx-Int Integration version of pMEND-Lx
pKW08-Lx TetRO promoter from pMEND-Lx inserted into the
backbone of pSE100; episomal
pKW08-Lx-Int Integration version of pKW08-Lx

Additional information

Can be purchased for academic use through Addgene: https://www.addgene.org/browse/article/3409/.

Publication

Williams KJ, Joyce G, Robertson BD. Improved mycobacterial tetracycline inducible vectors. Plasmid. 2010 Apr 29.

Inventor information

Dr Brian Robertson, Reader in Systems Microbiology, Department of Infectious Disease, Faculty of Medicine

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contact

Alexandra Skeaping

Industry Partnerships and Commercialisation Officer, Medicine

a.skeaping@imperial.ac.uk

+44 (0)20 7594 7021

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