Method for producing ceramide in transformed cells using the...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/00 (2006.01) C12N 9/02 (2006.01) C12N 15/53 (2006.01) C12N 15/81 (2006.01) C12P 13/02 (2006.01) C12P 7/64 (2006.01)

Patent

CA 2682712

Disclosed is a method for producing human ceramide in a cell such as a yeast cell. The method comprises the following steps 1) and 2): 1) introducing a gene (DES1m) for sphingolipid .DELTA.4-desaturase having an endoplasmic reticulum localization signal attached thereto by the transformation of a yeast cell, wherein the gene (DES1m) comprises sphingolipid .DELTA.4-desaturase gene (DES1) having a nucleotide sequence encoding a yeast endoplasmic reticulum localization signal attached to its 3'-terminus; and 2) causing a loss of the expression of yeast sphinganine C4-hydroxylase gene (SUR2) by the transformation of a yeast cell.

L'invention porte sur un procédé de fabrication de céramide humain dans une cellule telle qu'une cellule de levure. Le procédé comprend les étapes 1) et 2) suivantes : 1) introduire un gène (DES1m) pour la sphingolipide .DELTA.4-désaturase ayant un signal de localisation du réticulum endoplasmique fixé à celui-ci par la transformation d'une cellule de levure, le gène (DES1m) comprenant un gène de sphingolipide .DELTA.4-désaturase (DES1) ayant une séquence nucléotidique codant un signal de localisation du réticulum endoplasmique de levure fixé à son extrémité terminale 3' ; et 2) provoquer une perte de l'expression du gène de sphinganine C4-hydroxylase de levure (SUR2) par la transformation d'une cellule de levure.

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