Methods for the preparation of chemically misaminoacylated...

Details Methods for the preparation of chemically misaminoacylated... Methods for the preparation of chemically misaminoacylated...

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 21/00 (2006.01) C07H 21/02 (2006.01) C09K 11/07 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2473784

The present invention relates to methods for preparation of chemically aminoacylated tRNAs for the purpose of introduction of markers into nascent proteins. The present invention also relates to methods for the non- radioactive labeling, detection, quantitation and isolation of nascent proteins translated in a cellular or cell-free translation system utilizing chemically aminoacylated tRNAs. tRNA molecules are misaminoacylated with non- radioactive markers which may be non-native amino acids, amino acid analogs or derivatives. Markers may comprise cleavable moieties, detectable labels, reporter properties wherein markers incorporated into protein can be distinguished from unincorporated markers, or coupling agents which facilitate the detection and isolation of nascent protein from other components of the translation system.

L'invention porte: sur des procédés de préparation d'ARNt chimiquement aminoacylés servant à introduire des marqueurs dans des protéines naissantes; sur des procédés de marquage non radioactif, de détection, de quantification et d'isolement de protéines naissantes traduites dans un système cellulaire ou exempt de cellules à l'aide d'ARNt chimiquement aminoacylés. Les molécules d'ARNt sont mésaminoacylés à l'aide de marqueurs non radioactifs pouvant être des acides aminés non radioactifs, ou des analogues ou dérivés d'acides aminés. Les marqueurs peuvent consister en fragments clivables, indicateurs détectables aux propriétés de reporter lorsque les marqueurs incorporés à la protéine peuvent être distingués des marqueurs non incorporés, ou agents de couplage facilitant la détection et l'isolement de protéines naissantes parmi les autres constituants du système de traduction.

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