An electrophoretic method for the purification of RNA regions involved in protein crosslinking.
Direct information about structural interactions in ribonucleoprotein complexes can be obtained from crosslinking data. The purification of specific complexes, i.e., their separation from noncrosslinked proteins, from free RNA, and from other complexes, is essential for the identification of the bound proteins and the precise localization of their attachment sites in RNA. We describe a two-dimensional denaturing gel system which achieves this purification; in the first dimension basic proteins do not enter the gel and RNA--protein complexes are slowed down compared to protein free RNA, and in the second dimension sodium dodecyl sulfate improves the separation between the different complexes on the basis of their protein content.
Groupe de Photobiologie Moléculaire, Institut Jacques Monod, CNRS-Université Paris, France.
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Hajnsdorf
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Expert-Bezançon
D001426:Bacterial Proteins / Q000032:analysis; D005591:Chemical Fractionation; D015342:DNA Probes; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004926:Escherichia coli / Q000032:analysis; D007457:Iodine Radioisotopes; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012335:RNA, Ribosomal / Q000302:isolation & purification*; D012336:RNA, Ribosomal, 16S / Q000302:isolation & purification*; D012261:Ribonucleoproteins / Q000302:isolation & purification; D012269:Ribosomal Proteins / Q000302:isolation & purification
D001426:Bacterial Proteins; D015342:DNA Probes; D007457:Iodine Radioisotopes; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins; C040469:ribosomal protein S30