Skip to main content
. 2012 Apr 13;31(11):2579–2589. doi: 10.1038/emboj.2012.85

Figure 1.

Figure 1

Ribosomes containing MS2-tagged rRNAs can be affinity purified with GST–MS2. (A) Schematic representation of the plasmids used for rRNA expression. An 18-nt tag or MS2 tag was inserted into the 5′ region of the 25S rRNA. Both rRNAs were transcribed from the GAL7 promoter by RNA polymerase II. (B) Complementation test of an RNA polymerase I temperature-sensitive (ts) strain. The NOY401 strain containing various plasmids were grown and spotted onto SD–galactose plates after a series of 10-fold dilutions. (C–E) Protein and RNA compositions of affinity-purified ribosomes. (C) Each ribosomal fraction was isolated by sucrose density gradient sedimentation from the strain expressing the indicated plasmids and subjected to GST pull-down. The proteins were visualized by silver staining. Lanes 4, 8, and 12 show the intact ribosomal particles immunopurified with Rpl28–Flag from the fractions. GST–MS2cp: GST–MS2 coat protein. (D, E) RNAs were separated on the indicated gels and visualized with SYBR Gold staining. Figure source data can be found with the Supplementary data.