C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01) C12Q 1/00 (2006.01) C12Q 1/48 (2006.01) G01N 21/64 (2006.01) G01N 21/78 (2006.01) G01N 33/542 (2006.01) G01N 33/573 (2006.01) G01N 33/58 (2006.01)
Patent
CA 2410460
A widely applicable method of detecting and quantifying the phosphorylation/dephosphorylation of a protein in cells whereby the phosphorylation/dephosphorylation of a protein in vital cells, animals, plants, etc. can be non-destructively monitored and spatial and time-course analyses can be performed. In this method, use is made of a probe for visualizing phosphorylation/dephosphorylation characterized in that a tandem fusion unit, which has a substrate domain having a phosphorylation/dephosphorylation site and a phosphorylation-recognizing domain attached thereto via a linker sequence, is located between a donor chromophore and an acceptor chromophore in fluorescence resonance energy transfer.
L'invention concerne un procédé largement applicable de détection et de quantification de la phosphorylation/déphosphorylation d'une protéine dans des cellules. La phosphorylation/déphosphorylation d'une protéine dans des cellules vitales, des animaux, des végétaux, etc., peut être surveillée de manière non destructive et des analyses spatiales et temporelles peuvent être réalisées. Lors de ce procédé, on utilise une sonde pour visualiser la phosphorylation/déphosphorylation se caractérisant en ce qu'une unité de fusion en tandem, qui a un domaine de substrat doté d'un site de phosphorylation/déphosphorylation et un domaine de reconnaissance de la phosphorylation qui lui est fixé par le biais d'une séquence de liaison, est située entre un chromophore donneur et un chromophore accepteur en transfert d'énergie de résonance de la fluorescence.
Ozawa Takeaki
Sato Moritoshi
Umezawa Yoshio
Japan Science And Technology Corporation
Ogilvy Renault Llp/s.e.n.c.r.l.,s.r.l.
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