Protein synthesis monitoring (psm)

Details Protein synthesis monitoring (psm) Protein synthesis monitoring (psm)

C - Chemistry, Metallurgy

12

Q

C12Q 1/02 (2006.01) C12M 1/00 (2006.01) C12Q 1/00 (2006.01) C12Q 1/34 (2006.01) C12Q 1/37 (2006.01) C12Q 1/48 (2006.01) G01N 33/53 (2006.01)

Patent

CA 2517566

A method and a device are disclosed for monitoring the synthesis of proteins by the ribosome in real time, in-vivo as well as in in-vitro. The ribosome is engineered to carry a donor fluorophore, and tRNA and/or amino acids and/or some other part of the ribosome are either engineered to carry acceptor fluorophores or else their natural fluorescent properties are utilized as acceptors. As the ribosome mechanism processed the mRNA and tRNA molecules and synthesizes a polypeptide chain, a light source illuminates the ribosome, exciting the donor fluorophores and thereby the acceptor fluorophores whenever these are in sufficient proximity to a donor. The resulting signals are detected and used as a key for real-time database searching and identification of the protein being synthesized. The resulting data can be tabulated and interpreted in different ways. Figure (1) describes the propertires of a FRET pair and the dependence of FRET on pair distance.

Cette invention concerne un procédé et un dispositif permettant de surveiller la protéinogenèse par le ribosome, en temps réel,in vivo et in vitro. Le ribosome est conçu pour porte un fluorophore donneur. L'ARNt et/ou des acides aminés et/ou certaines autres parties du ribosome sont conçus pour porter des fluorophores accepteurs, ou bien encore leur propriétés fluorescentes naturelles sont utilisées comme accepteurs. Lorsque le mécanisme ribosomique transforme les molécules d'ARNm et d'ARNt et synthétise une chaîne polypeptidique, une source lumineuse éclaire le ribosome, ce qui excite les fluorophores donneurs et, par là même, les fluorophores accepteurs lorsqu'ils sont suffisamment proches d'un donneur. Les données résultantes peuvent être totalisées, le but étant d'interpréter l'activité de médicaments candidats, d'élucider des mécanismes protéomiques cellulaires, de gérer la production de protéines et de façon générale, de recueillir des informations nouvelles et importantes déterminantes pour la résolution de l'énigme cellulaire.

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