System for screening cells for high expression of a protein...

C - Chemistry – Metallurgy – 07 – K

Patent

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Details

C07K 19/00 (2006.01) C12N 5/10 (2006.01) C12N 15/62 (2006.01) C12N 15/63 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2618203

This invention refers to industrial production of proteins. More particularly, the invention refers to a fusion protein as a novel chimeric selection marker comprising a peptide conferring resistance to an antibiotic, or a fragment, allelic variant, splice variant or mutein thereof, and at least one sequence comprising SEQ ID NO: 1, 2 or 3, preferably for producing a protein of interest (POI). The inventive chimeric selection marker exhibits: (i) a resistance to an antibiotic; and (ii) a fluorescence activity upon binding of a ligand to the sequence comprising SEQ ID NO: 1, 2 or 3. The invention further refers to nucleic acids encoding the inventive fusion protein and to expression vectors, comprising the inventive fusion protein and additionally the protein of interest (POI). Finally, uses of the inventive chimeric selection marker for screening cells for high expression of a protein of interest (POI) are disclosed.

L'invention concerne la production industrielle de protéines. Elle concerne notamment une protéine de fusion utilisée comme un nouveau marqueur de sélection chimérique comprenant un peptide conférant la résistance à un antibiotique, ou leur fragment, variant allélique, variant épissé variant ou mutéine et au moins une séquence comprenant SEQ ID NO: 1, 2 ou 3, de préférence pour la production d'une protéine d'intérêt (POI). Le marqueur de sélection chimérique de l'invention manifeste (i) une résistance à un antibiotique; et (ii) une activité de fluorescence après la liaison d'un ligand à la séquence comprenant SEQ ID NO: 1, 2 ou 3. L'invention concerne également des acides nucléiques codant la protéine de fusion de l'invention et des vecteurs d'expression, y compris la protéine de fusion de l'invention et, en outre, la protéine d'intérêt (POI). Finalement, l'invention concerne des utilisations du marqueur de sélection chimérique de l'invention pour cribler des cellules destinées à l'expression élevée d'une protéine d'intérêt (POI).

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