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Characterization of Hairpin Ribozyme Reactions

  • Preeti Bajaj
  • Christian Hammann
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1103)

Abstract

Hairpin ribozymes are small RNA catalytic motifs naturally found in the satellite RNAs of tobacco ringspot virus (TRsV), chicory yellow mottle virus (CYMoV), and arabis mosaic virus (ArMV). The catalytic activity of the hairpin ribozyme extends to both cleavage and ligation reactions. Here we describe methods for the kinetic analysis of the self-cleavage reaction under transcription reaction conditions. We also describe methods for the generation of DNA templates for subsequent in vitro transcription reaction of hairpin ribozymes. This is followed by a description of the preparation of the suitable RNA molecules for ligation reaction and their kinetic analysis.

Key words

Catalytic RNA Kinetic analysis Plant virus satellite RNA Self-cleavage Ligation 

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Preeti Bajaj
    • 1
  • Christian Hammann
    • 2
  1. 1.Department of Insect ResistanceInternational Center for Genetic Engineering and BiotechnologyNew DelhiIndia
  2. 2.Ribogenetics @ Biochemistry Lab, Molecular Life Sciences Research Center, School of Engineering and ScienceJacobs UniversityBremenGermany

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