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. 2017 Apr 11;45(11):6837–6847. doi: 10.1093/nar/gkx254

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 1. — Purification of plasmid-derived pre-60S particles. (A) Construction of pre-27S RNA fragments. The structural diagram and processing sites of 35S pre-rRNA are shown on the top. Seven pre-27S RNA fragments terminate after one of six domains I–VI (D1–D6) of 25S rRNA or the 3΄ ETS and contain an MS2-tag (square) in ITS2 and a plasmid-specific sequence (circle, 25S-tag) in domain I. These RNAs are transcribed under a GAL-promoter in multiple-copy 2μ plasmids. Locations of hybridization probes are indicated. (B) Plasmid-derived pre-60S particles were affinity purified via an MS2-tag in pre-rRNA and then via TAP-tagged Nop7.