UNIT 27.3 Purification of Functional RNA-Protein Complexes using MS2-MBP
Published Online: 1 AUG 2003
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Zhou, Z. and Reed, R. 2003. Purification of Functional RNA-Protein Complexes using MS2-MBP. Current Protocols in Molecular Biology. 63:27.3:27.3.1–27.3.7.
- Published Online: 1 AUG 2003
- Published Print: JUL 2003
Biological machines composed of RNAs and proteins play essential roles in many biological processes. To better understand the mechanism and function of these machines, it is critical to isolate them in a highly purified and functional form. A method for isolating functional RNA-protein complexes assembled in vitro is described. The approach combines gel filtration and an affinity-chromatography strategy using the bacteriophage MS2 coat protein, which binds to a specific RNA-hairpin structure. Using this method, highly purified and functional human spliceosomes have been isolated. The purified spliceosome preparation is used to determine the protein components of the spliceosome by mass spectrometry and to examine the structure of the spliceosome by electron microscopy.
- RNA-protein complex;
- ribonucleoprotein complex;
- affinity purification