Chitin synthase 1

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/54 (2006.01) C07K 16/40 (2006.01) C12N 9/10 (2006.01) C12N 15/81 (2006.01) C12Q 1/02 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2233881

A polynucleotide encoding chitin synthase (CHS1), an enzyme essential for cell wall synthesis and yeast cell growth, is provided. A maltose responsive promoter (MRP) isolated using the promoter library of the invention is also described. The present invention also provides a vector for isolation of a eukaryotic regulatory polynucleotide, i.e., promoter. The vector is useful in the method of the invention which comprises identifying a eukaryotic regulatory polynucleotide, i.e., promoter region, by complementing the growth of an auxotrophic host cell containing the vector of the invention, which includes a promoter region operably linked to a promoterless auxotrophic gene. The vector is introduced into the host cell chromosome by targeted integration. Also provided is a library containing host cells having the vector of the invention integrated in the chromosome of the host cell.

L'invention porte sur un polynucléotide codant pour la chitine synthase 1, enzyme essentielle à la synthèse de la paroi cellulaire et à la croissance des cellules de levure. L'invention porte également sur un promoteur eucaryote réagissant au maltose, isolé à l'aide d'une bibliothèque de promoteurs, objet de l'invention. Elle porte en outre sur un vecteur d'isolement d'un polynucléotide régulateur eucaryote (le promoteur). Ledit vecteur est utile dans le procédé objet de l'invention consistant à identifier la région du polynucléotide régulateur eucariote (le promoteur) pour compléter la croissance d'une cellule hôte auxotrophique contenant le vecteur objet de l'invention et qui comporte une région promotrice fonctionnellement liée à un gène auxotrophique sans promoteur. Le vecteur est introduit dans le chromosome de la cellule hôte par intégration ciblée. L'invention porte enfin sur une bibliothèque de cellules hôtes au chromosome desquelles le vecteur objet de l'invention est intégré.

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