Expression system for proteins

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/67 (2006.01)

Patent

CA 2716905

The invention provides a novel system for the tunable expression of nucleic acids encoding e.g., polypeptides such as recombinant proteins in prokaryotic systems. The system is based on the ability of T7 lysozyme (T7Lys) to inhibit the activity of T7R-NAP. Expression of T7Lys can be continuously adjusted as its expression is under the control of a promoter whose activity can be titrated. The invention provides a host cell capable of expressing T7 RNA polymerase, the host cell comprising a first nucleic acid having a T7 lysozyme gene or a T7 lysozyme variant gene and a tunable promoter for controlling the expression of the T7 lysozyme gene. It also provides a host cell further comprising a second nucleic acid having a T7 promoter operably linked to a nucleic acid sequence encoding a target polypeptide, whereby expression of the target polypeptide is tuned via controlling the expression of the T7 lysozyme gene.

La présente invention concerne un nouveau système destiné à lexpression modulable dacides nucléiques codant notamment pour des polypeptides tels que des protéines recombinantes dans des systèmes procaryotes. Le système est fondé sur la capacité du lysozyme T7 (T7Lys) dinhiber lactivité de lARN polymérase T7 (T7RNAP). Lexpression de T7Lys peut être sans cesse ajustée lorsque son expression est sous la régulation dun promoteur dont lactivité peut être dosée. Linvention concerne une cellule hôte capable dexprimer lARN polymérase T7, la cellule hôte étant pourvue dun premier acide nucléique qui comporte un gène du lysozyme T7 ou un variant de gène du lysozyme T7 et un promoteur pouvant être modulé pour réguler lexpression du gène du lysozyme T7. Linvention concerne également une cellule hôte pourvue en outre dun second acide nucléique qui comporte un promoteur de T7 fonctionnellement lié à une séquence d'acide nucléique codant pour un polypeptide cible, par quoi lexpression du polypeptide cible est modulée par la régulation de lexpression du gène du lysozyme T7.

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