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US 2003/0190643 A1Published application
Sequencing by mass spectrometry
AssigneeTrustees of Boston University
InventorsKenneth J. Rothschild, Sadanand Gite, Jerzy Olejnik
Life of this patent
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Abstract
This invention relates to non-radioactive markers that facilitate the detection and analysis of nascent proteins translated within cellular or cell-free translation systems. Nascent proteins containing these markers can be rapidly and efficiently detected, isolated and analyzed without the handling and disposal problems associated with radioactive reagents. Preferred markers are dipyrrometheneboron difluoride (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene)dyes.
Claims20
A method, comprising:
- a) providing a tRNA molecule and a mass marker;
- b) aminoacylating said tRNA molecule with said mass marker to create a misaminoacylated tRNA;
- c) introducing said misaminoacylated tRNA into a translation system under conditions such that said marker is incorporated into a nascent protein;
- and
- d) detecting said mass marker by mass spectrometry.
2 more elements
A method, comprising:
- a) providing a tRNA molecule and a mass marker;
- b) aminoacylating said tRNA molecule with said mass marker to create a misaminoacylated tRNA;
- c) introducing said misaminoacylated tRNA into a translation system under conditions such that said marker is incorporated into a nascent protein;
- d) proteolyzing said nascent protein;
- and
- e) detecting said mass marker by mass spectrometry.
3 more elements
A method, comprising:
- a) providing a tRNA molecule and a photocleavable mass marker;
- b) aminoacylating said tRNA molecule with said photocleavable mass marker to create a misaminoacylated tRNA;
- c) introducing said misaminoacylated tRNA into a translation system under conditions such that said photocleavable mass marker is incorporated into a nascent protein;
- and
- d) exposing said photocleavable mass marker to electromagnetic radiation.
2 more elements
Dependencies are inferred from the claim text.