D-amino acid oxidase promoter from t. variabilis

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 1/19 (2006.01) C07H 21/04 (2006.01) C07K 14/495 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 9/06 (2006.01) C12N 9/18 (2006.01) C12N 9/86 (2006.01) C12N 15/81 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2292844

A D-amino acid oxidases promoter isolated from T. variabilis has been incorporated into S. cerevisiae expression vectors to increase the expression of heterologous genes. The heterologous promoter has been placed upstream of selective markers, auxotrophic nutritional and/or dominant selectable markers. Because the promoter is from another yeast specie, it is weakly recognized by S. cerevisiae. To compensate for the weak transcripts of selectable marker genes transcribed from the weak heterologous promoter, the plasmid containing the gene of the interest is amplified leading to a higher expression of protein of interest.

On incorpore un promoteur de D-amino acide oxydase extrait de T. variabilis dans des vecteurs d'expression de S. cerevisiae pour augmenter l'expression de gènes hétérologues. On place le promoteur hétérologue en amont de marqueurs sélectifs, de marqueurs nutritionnels auxotrophes et/ou de marqueurs dominants pouvant être sélectionnés. Etant donné que le promoteur est tiré d'une autre espèce de levure, il n'est que faiblement reconnu par S. cerevisiae. Afin de compenser la faible transcription des gènes marqueurs pouvant être sélectionnés transcrits à partir du promoteur hétérologue faible, le plasmide contenant le gêne étudié est amplifié, ce qui entraîne un degré d'expression plus élevé de la protéine considérée.

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