Two green fluorescent protein fragments and their use in a...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/12 (2006.01) C07K 14/435 (2006.01) G01N 33/542 (2006.01) G01N 33/58 (2006.01) G01N 33/68 (2006.01)

Patent

CA 2482897

Fluorescence complementation products with intensity levels mimicking the full length intensities are obtained by introduction of improved folding capabilities with a mutation in position preceding the chromophore. This is particularly seen with the yellow variant of Green Fluorescent Protein (GFP). An additive increase is obtained by splitting the GFP between amino acids 172 and 173. Screening for drugs capable of preventing interaction between proteins is performed by selecting the cells with the highest dynamic range through Fluorescence Activated Cell Sorting (FACS), as illustrated with the ability of FK506 to break the rapamycin induced interaction between FRB and FKBP.

On obtient des produits de complémentation de fluorescence à niveaux d'intensité imitant des intensités pleine longueur par introduction de capacités de pliage amélioré avec une mutation en position 1 précédant le chromophore. On le remarque surtout avec la variante jaune de la protéine verte fluorescente (GFP). On obtient une augmentation de l'additif par séparation de la protéine GFP en acides aminés (172 et 173). On effectue le criblage de médicaments permettant de prévenir l'interaction entre les protéines par sélection de cellules possédant la plage dynamique la plus élevée au moyen d'un trieur de cellules à fluorescence (FACS), tel qu'illustré avec la capacité de FK506 à rompre l'interaction induite par la rapamycine entre FRB et FKBP.

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