C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/04 (2006.01) C07K 19/00 (2006.01) C12M 1/34 (2006.01) C12N 5/10 (2006.01) C12N 15/63 (2006.01) C12Q 1/00 (2006.01) C12Q 1/02 (2006.01) G01N 21/64 (2006.01) G01N 33/50 (2006.01) G01N 33/68 (2006.01)
Patent
CA 2444857
The present invention relates to a two-hybrid system for studying protein- protein interactions in mammalian host cells. The invention provides methods, reagents and kits for carrying out the two-hybrid screen. Accordingly the invention provides a bait construct that is capable of targeting a bait protein to a specific subcellular locale within the host cell. The invention also provides a prey construct that contains a detection sequence fused to a prey protein. The bait and prey constructs are introduced into a host cell under conditions that promote expression of the bait and prey constructs. A positive bait/prey interaction can be detected by comparing the subcellular localization of the prey construct in relation to the bait construct. The invention also provides methods for screening for compounds capable of disrupting protein-protein interaction and methods for detecting interactions between proteins and small molecule compounds.
La présente invention concerne un système à deux hybrides permettant d'étudier les interactions entre protéines dans des cellules hôtes mammaliennes. L'invention propose des procédés, des réactifs et des trousses permettant d'effectuer un criblage à deux hybrides. A cet effet, l'invention propose une construction d'appât susceptible de cibler une protéine appât vers un site infracellulaire au sein d'une cellule hôte. L'invention propose également une construction de proie qui contient une séquence de détection fusionnée à la protéine proie. Les constructions d'appât et de proie sont insérées dans la cellule hôte dans des conditions pour favoriser l'expression des constructions d'appât et de proie. On peut détecter une interaction appât/proie en comparant la localisation infracellulaire de la construction de proie par rapport à la construction d'appât. L'invention concerne en outre des procédés permettant le criblage de composés capables d'interrompre l'interaction entre protéines et des procédés de détection des interactions entre des protéines et de composés à petites molécules.
Mckeon Frank
Yang Annie
Bereskin & Parr Llp/s.e.n.c.r.l.,s.r.l.
President And Fellows Of Harvard College
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