Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 May 10;111(1):304–315. doi: 10.1111/tpj.15779

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2022 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

PMC Copyright notice

Schematic of the translating ribosome affinity purification (TRAP) workflow.

(a) Tissue from transiently transformed Nicotiana benthamiana or stably transformed Arabidopsis was collected, flash‐frozen, and homogenized with extraction buffer. Clarification of the homogenate via centrifugation yielded the clarified leaf extract (CLE), containing all soluble proteins.

(b) Polysome concentration was achieved by ultracentrifugation of CLE over a sucrose cushion followed by resuspension, yielding the sucrose cushion ultracentrifugation pellet (SUP) containing all protein complexes with sufficient density to penetrate the cushion. Note that this step was omitted from the experiments with transgenic Arabidopsis. (c) Immunopurification of tagged polyribosomes from either CLE (Arabidopsis) or SUP (N. benthamiana) employed FLAG beads to yield immunoprecipitated (IP) polysomes. [Colour figure can be viewed at wileyonlinelibrary.com]