Methods using o6-alkylguanine-dna alkyltransferases

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/48 (2006.01) G01N 33/531 (2006.01)

Patent

CA 2443570

A method using O6-alkylguanine-DNA alkyltransferases (AGT) is disclosed for transferring a label from a substrate to a fusion protein comprising the AGT. This allows the detection and/or manipulating of the fusion protein, both in vitro and in vivo, by attaching molecules to the fusion proteins that introduce a new physical or chemical property to the fusion protein. Examples of such molecules are, among others, spectroscopic probes or reporter molecules, affinity tags, molecules generating reactive radicals, cross- linkers, ligands mediating protein-protein interactions or molecules suitable for the immobilisation of the fusion protein.

L'invention porte sur un procédé d'utilisation de O6-alkylguanine-DNA alkyltransférase (AGT) permettant de transférer un marqueur depuis un substrat vers une protéine de fusion comprenant l'AGT. Ce procédé permet de détecter et/ou de manipuler la protéine de fusion, à la fois in vitro et in vivo, en attachant des molécules aux protéines de fusion qui introduisent une nouvelle propriété physique ou chimique dans la protéine de fusion. Parmi les exemples de ces molécules on trouve, entre autres, des sondes spectroscopiques ou des molécules <= reporters >=, des marqueurs d'affinité, des molécules générant des radicaux réactifs, des agents de réticulation, des ligands qui induisent des interactions protéines-protéines ou des molécules qui servent à immobiliser la protéine de fusion.

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