Insect-derived promoters for foreign proteins expression in...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/85 (2006.01) C12N 15/866 (2006.01) A01K 67/033 (2006.01)

Patent

CA 2735259

Insect-derived promoters for foreign proteins expression in insect cells. Regulatory polynucleotide sequences which drive the expression of major insect proteins (hexamerins) at specific evolution stages of larva have been isolated from insects (Trichoplusia ni) in the present invention. Said regulatory polynucleotide sequences promote stronger foreign gene expression in the baculovirus system than the conventional polyhedrin promoter. Additionally, the combination of the new larva-derived promoters of the invention with the pL promoter increased baculovirus expression levels up to 61%-375% (depending on the time of infection) with respect to conventional baculo viruses used in the biotechnology industry. Promoter pB2 also drives gene ex-pression at earlier times than polyhedrin promoter, being this feature a great advantage for correct protein folding and posttranslational modification.

L'invention concerne des promoteurs dérivés d'insectes pour l'expression de protéines étrangères dans des cellules d'insectes. Des séquences polynucléotidiques régulatrices commandant l'expression de protéines d'insectes majeures (hexamérines) à des étapes d'évolution spécifiques de larves ont été isolées d'insectes (Trichoplusia ni). Lesdites séquences polynucléotidiques régulatrices promeuvent une expression plus forte de gènes étrangers dans le système baculoviral par rapport au promoteur polyhédrine habituel. Par ailleurs, la combinaison de nouveaux promoteurs dérivés de larves au promoteur pL permet d'augmenter les niveaux d'expression du baculovirus de l'ordre de 61 à 375 % (en fonction de la durée de l'infection) par rapport aux baculovirus habituels employés en biotechnologie. Le promoteur pB2 commande également l'expression génétique de façon plus précoce que ne le fait le promoteur polyhédrine, cette caractéristique constituant un avantage considérable pour le repliement et les modifications post-translationnelles des protéines.

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