Fluorescent labeling of transfer RNA and study of protein synthesis
US9612244B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Jun 3, 2014 |
| Grant date | Apr 4, 2017 |
| Priority date | — |
| Expiry date | Oct 4, 2034 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/6803
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Provided are methods for labeling transfer RNA comprising replacing the uracil component of a dihydrouridine of said transfer RNA with a fluorophore. The disclosed methods may comprise fluorescent labeling of natural tRNAs (i.e., tRNAs that have been synthesized in a cell, for example, in a bacterium, a yeast cell, or a vertebrate cell) at dihydrouridine (D) positions, or fluorescent labeling of synthetic tRNAs. In another aspect, the present invention provides methods for assessing protein synthesis in a translation system comprise providing a tRNA having a fluorophore substitution for the uracil component of a dihydrouridine in a D loop of the tRNA; introducing the labeled tRNA into the translation system; irradiating the translation system with electromagnetic radiation, thereby generating a fluorescence signal from the fluorophore; detecting the fluorescence signal; and, correlating the fluorescence signal to one or more characteristics of the protein synthesis in the translation system. The disclosed methods are useful in single molecule as well as in ensemble settings.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.