A coumermycin/novobiocin-regulated gene expression system

C - Chemistry – Metallurgy – 12 – N

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C12N 15/67 (2006.01)

Patent

CA 2523879

A chimeric transactivator comprises a transcription activation domain, a repressor protein DNA binding domain and the bacterial DNA gyrase B subunit. A target gene is operatively linked to operator DNA sequences recognized by the repressor binding domain. The addition of the antibiotic coumermycin results in a coumermycin-switched dimerization of the transactivator, which then binds to operator DNA sequences and activates transcription of the target gene. The addition of novobiocin switches off expression of the target gene by abolishing coumermycin-induced dimerization of the transactivator.

L'invention concerne un transactivateur chimérique comprenant un domaine d'activation de transcription, un domaine de liaison à l'ADN d'une protéine répresseur et la sous-unité B de l'ADN gyrase bactérienne. Un gène cible est lié fonctionnel aux séquences ADN opérateur reconnues par le domaine de liaison du répresseur. L'addition de la coumermycine antibiotique entraîne une dimérisation activée par la coumermycine du transactivateur, qui se lie ensuite aux séquences ADN opérateur et active la transcription du gène cible. L'addition de la novobiocine désactive l'expression du gène cible par élimination de la dimérisation induite par la coumermycine du transactivateur.

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