Emission ratiometric indicators of phosphorylation

Details Emission ratiometric indicators of phosphorylation Emission ratiometric indicators of phosphorylation

C - Chemistry, Metallurgy

12

N

C12N 15/62 (2006.01) C12N 15/00 (2006.01) C12N 15/63 (2006.01) C12Q 1/42 (2006.01) C12Q 1/48 (2006.01) G01N 33/52 (2006.01) G01N 33/53 (2006.01) G01N 33/542 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2447799

A chimeric phosphorylation indicator is provided. A chimeric phosphorylation indicator can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. A chimeric phosphorylation indicator also can contain a phosphorylatable polypeptide and a fluorescent protein, wherein the phosphorylatable polypeptide is contained within the sequence of the fluorescent protein, or wherein the fluorescent protein is contained within the sequence of the phosphorylatable polypeptide. Also provided are polynucleotides encoding such chimeric phosphorylation indicators, as well as kits containing the indicators or the polynucleotides. In addition, a method of using the chimeric phosphorylation indicators to detect a kinase or phosphatase in a sample is provided.

L'invention concerne un indicateur de phosphorylation chimérique. Un indicateur de phosphorylation chimérique peut contenir un molécule donneuse, un domaine phosphorylable, un domaine de liaison de phospho-amino acide (PAABD), et une molécule receveuse. Un indicateur de phosphorylation chimérique peut également contenir un polypeptide phosphorylable et une protéine fluorescente, le polypeptide phosphorylable étant contenu à l'intérieur de la séquence de la protéine fluorescente, ou la protéine fluorescente étant contenue à l'intérieur de la séquence du polypeptide phosphorylable. L'invention concerne également des polynucléotides codant de tels indicateurs de phosphorylation chimérique, ainsi que des trousses de diagnostic contenant lesdits indicateurs ou lesdits polynucléotides. En outre, un procédé d'utilisation des indicateurs de phosphorylation chimérique permet de détecter une kinase ou une phosphatase dans un échantillon.

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