Expression vector for producing protein derived from foreign...

Details Expression vector for producing protein derived from foreign... Expression vector for producing protein derived from foreign...

C - Chemistry, Metallurgy

12

N

C12N 15/09 (2006.01) C12P 21/02 (2006.01) C12Q 1/02 (2006.01)

Patent

CA 2748011

Studies have been made extensively, and such an expression vector can be successfully constructed that enables the high-level production of a protein derived from a foreign gene in a mammal host cell. The expression vector comprises a gene cassette which contains a translation-impairing dihydrofolate reductase gene cistron and a cloning site into which a foreign gene is to be integrated, wherein codons in the translation-impairing dihydrofolate reductase gene cistron are altered to those which have been used in mammal cells with lowest frequencies to reduce the expression level thereof, and the cloning site is located between a highly transcriptionally active promoter and a highly stable polyadenylation signal.

Des études ont été menées de manière extensive, et un tel vecteur d'expression peut être construit avec succès qui permet la production à un niveau élevé d'une protéine issue d'un gène étranger dans une cellule hôte de mammifère. Le vecteur d'expression comprend une cassette de gène qui contient un cistron de gène de dihydrofolate réductase affectant la traduction et un site de clonage dans lequel un gène étranger doit être intégré, des codons dans le cistron de gène de dihydrofolate réductase affectant la traduction étant altérés en ceux qui ont été utilisés dans des cellules de mammifère avec les fréquences les plus faibles afin de réduire le niveau d'expression de celui-ci, et le site de clonage est situé entre un promoteur à activité transcriptionnelle élevée et un signal de polyadénylation hautement stable.

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