snRNPs: Methods for Purification

Berthold Kastner, Phillipps University of Marburg, Marburg, Germany

Published online: April 2001

DOI: 10.1038/npg.els.0002605

Small nuclear ribonucleoprotein particles (snRNPs) are essential cofactors for the nuclear splicing of pre-mRNA introns. The method best suited to the purification of snRNPs is affinity chromatography combined with competitive elution.

Keywords: snRNP; pre-mRNA splicing; chromatography; RNA–protein interaction

Figure 1. Glycerol gradient centrifugation of snRNPs. snRNPs eluted from an anti-m3G column were loaded on a 10–30% glycerol gradient and centrifuged in a Beckman SW 28 rotor for 16 h at 27 000 rpm. After fractionation the proteins were extracted and separated by electrophoresis on an SDS–polyacrylamide gel. The migration of the individual snRNAs is indicated at the bottom of the figure. The bands corresponding to the common proteins are marked on the left, and on the right the approximate molecular masses of several specific proteins are indicated. The Coomassie-stained gel was kindly provided by Axel Badouin.
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 References
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 Further Reading
    book Kastner B (1998) "Purification and electron microscopy of spliceosomal snRNPs". In: Schenkel J (ed.) RNP. Particles, Splicing and Autoimmune Diseases, pp. 95–140. Berlin: Springer Verlag.
    Krämer A (1990) Purification of small nuclear ribonucleoprotein particles active in RNA processing. Methods in Enzymology 181: 215–231.
    book Will CL, Kastner B and Lührmann R (1994) "Analysis of ribonucleoprotein interactions". In: Higgens SJ and Hames BD (eds) RNA Processing. – A Practical Approach, vol. 1, pp. 141–177. Oxford: IRL Press.

Related Sub-Topics:

Transcription | Sample Preparation in Structural Biology | Techniques and Tools in Clinical Genetics | Proteomic Studies of Genetic Disease | Transcription of DNA | RNA Structure and Function | Techniques and Tools in Molecular Biology
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Article Title: snRNPs: Methods for Purification
 
How to Cite close
Kastner, Berthold(Apr 2001) snRNPs: Methods for Purification. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002605]