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Isolation of homogenous ribosome-nascent-chain complexes from coupled cell-free transcription-translation system

External protocol Created on 30 Apr 2014

Authors

Gong Zhang and Zoya Ignatova

Summary

Recent studies have made some progress in tracking the conformation of the nascent chains1-3; however the earliest steps of folding in vivo (while chain synthesis is underway) remain unclear. This is in part due to a lack of experimental methods to produce stable ribosome-nascent chain complexes with uniform length of the nascent chains. Translation mixtures usually contain heterogeneous fraction of ribosomes with various lengths of the peptide chains due to re-initiation of the translation; this hinders the precise assessment of the conformation of the nascent chains. Here, we describe a detailed procedure to produce homogeneous fractions of ribosome-nascent-chain complexes with defined length of the stalled peptides.

Further details

The protocol was published on Protocol Exchange on 16 February 2009. To see the entire protocol, click on the source link.

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