Steroid receptor modulation of gene expression

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 1/15 (2006.01) C12N 1/19 (2006.01) C12N 15/63 (2006.01) C12N 15/80 (2006.01) C12N 15/81 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2511359

The present invention provides a novel steroid inducible expression system in a non-mammalian host cell (e.g., fungal) that is independent of metabolic and development regulation. The human estrogen receptor gene expressed in Aspergillus, under a constitutive promoter, was shown to be functional. A reporter gene containing regulatory sequences from Aspergillus, yeast and a synthetic sequence containing the estrogen receptor binding sites (EREs) was expressed in response to a hormone derivative inducer. In the absence of the inducer, the promoter is silent and depending on the type of construct and inducer concentration the expression level can e modulated from moderate to very strong.

L'invention concerne un nouveau système d'expression inductible par des stéroïdes dans une cellule hôte non mammalien (par exemple fongique) indépendant de la régulation métabolique et du développement. Le gène du récepteur de l'oestrogène humain exprimé dans l'Aspergillus, sous un promoteur constitutif, s'est avéré fonctionnel. Un gène rapporteur contenant des séquences régulatrices de l'Aspergillus, une levure et une séquence synthétique contenant les sites de liaison du récepteur de l'oestrogène (ERE), est exprimé en réponse à un inducteur dérivé hormonal. En l'absence de l'inducteur, le promoteur est silencieux et, selon le type de construction et la teneur de l'inducteur, le niveau d'expression peut être modulé de modéré à très fort.

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