Real-Time Characterization of Ribozymes by Fluorecence Resonance Energy Transfer (FRET)

Andreas Jenne; Walter Gmelin; Nikolai Raffler; Michael Famulok

doi:10.1002/(SICI)1521-3773(19990503)38:9<1300::AID-ANIE1300>3.0.CO;2-Q

Real-Time Characterization of Ribozymes by Fluorecence Resonance Energy Transfer (FRET)

Angew Chem Int Ed Engl. 1999 May 3;38(9):1300-1303. doi: 10.1002/(SICI)1521-3773(19990503)38:9<1300::AID-ANIE1300>3.0.CO;2-Q.

Authors

Andreas Jenne¹, Walter Gmelin¹, Nikolai Raffler¹, Michael Famulok¹

Affiliation

¹ Institut für Biochemie der Universität, Feodor-Lynen-Strasse 25, D-81377 Munich (Germany), Fax: (+49) 89-74017-448.

Abstract

More rapid than with conventional methods is the analysis of ribozyme kinetics upon use of FRET substrates. In these substrates a fluorophore and a fluorescence-quenching molecule lie in close spatial proximity. The intramolecular fluorescence quenching is neutralized upon cleavage, and the fluorescence intensity is a measure of the ribozyme activity. By automation and computer assistance the activity of ribozymes can be monitored under high-throughput conditions.

Keywords: Fluorescence spectroscopy; Hammerhead ribozyme; Ribozymes; Screening methods.