Fluorescent protein sensors for detection of analytes

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C07K 14/435 (2006.01) C12N 15/09 (2006.01) G01N 33/542 (2006.01)

Patent

CA 2283629

Fluorescent indicators including a binding protein moiety, a donor fluorescent protein moiety, and an acceptor fluorescent protein moiety are described. The binding protein moiety has an analyte-binding region which binds an analyte and causes the indicator to change conformation upon exposure to the analyte. The donor moiety and the acceptor moiety change position relative to each other when the analyte binds to the analyte-binding region. The donor moiety and the acceptor moiety exhibit fluorescence resonance energy transfer when the donor moiety is excited and the distance between the donor moiety and the acceptor moiety is small. The indicators can be used to measure analyte concentrations is samples, such as calcium ion concentrations in cells.

L'invention concerne des indicateurs fluorescents comprenant une fraction de protéine de liaison, une fraction de protéine fluorescente donneuse, et une fraction de protéine fluorescente accepteuse. La fraction de protéine de liaison comporte une région de liaison de substance à analyser qui se lie à une substance à analyser et provoque un changement de conformation de l'indicateur après exposition à la substance à analyser. La fraction donneuse et la fraction accepteuse changent de position l'une par rapport à l'autre lorsque la substance à analyser se lie à la région de liaison de substance à analyser. La fraction donneuse et la fraction accepteuse présentent un transfert d'énergie de résonance de fluorescence lorsque la fraction donneuse est excitée et que la distance entre la fraction donneuse et la fraction accepteuse est réduite. Les indicateurs peuvent s'utiliser pour mesurer des concentrations de substances à analyser dans des échantillons, telles que des concentrations d'ion calcium dans des cellules.

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