Recombinant expression of multiprotein complexes using...

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/79 (2006.01) A01K 67/027 (2006.01) A61K 48/00 (2006.01) C12N 15/10 (2006.01) C12N 15/866 (2006.01)

Patent

CA 2626891

The present invention relates to a recombinant polynucleotide encoding a polygene coding for at least three polypeptides wherein at least one of the genes constituting the polygene is of non-viral origin, at least two of the polypeptides encoded by the genes constituting the polygene are each capable of at least transiently interacting with at least one other polypeptide encoded by a gene of said polygene, and the genes constituting the polygene are each connected to one another by a sequence coding for at least one protease cleavage site. The present invention also relates to polyproteins encoded by the polygene. Further embodiments of the present invention are a vector containing the recombinant polypeptide, a host cell containing the recombinant polypeptide and/or the vector and a non-human transgenic animal transformed with the recombinant polypeptide and/or the vector. The present invention also relates to methods for the production of the polynucleotide and for the manufacture of multiprotein complexes. The embodiments of the present invention are particularly useful in gene therapy, drug candidate screening, vaccine production and crystallisation of multiprotein complexes for structural investigations.

L'invention concerne un polynucléotide recombiné qui code pour un polygène codant pour au moins trois polypeptides, au moins un des gènes constituant le polygène étant d'origine non virale, au moins deux des polypeptides codés par les gènes constituant le polygène étant chacun capables d'interagir au moins de manière au moins transitoire avec au moins un autre polypeptide codé par un gène dudit polygène; et les gènes constituant le polygène étant chacun reliés entre eux par une séquence codant pour au moins un site de coupure de protéase. L'invention concerne aussi des polyprotéines codées par le polygène. D'autres formes de réalisation de l'invention concernent un vecteur contenant le polypeptide recombiné, une cellule hôte contenant le polypeptide recombiné et/ou le vecteur, et un animal transgénique non humain transformé à l'aide du polypeptide recombiné et/ou du vecteur. L'invention concerne aussi des procédés de production du polynucléotide et des procédés de fabrication de complexes multiprotéiques. Les formes et modes de réalisation de l'invention sont particulièrement utiles en thérapie génique, dans le criblage de médicaments candidats, la production de vaccins et la cristallisation de complexes multiprotéiques à des fins d'études structurales.

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