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Ribosome Profiling

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...

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Updated: May 27, 2026

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes

Published on: October 3, 2019

Peptide screening using PURE ribosome display.

Hiroyuki Ohashi¹, Takashi Kanamori, Eriko Osada

¹Department of Medical Genome Sciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba Prefecture, Japan.

Methods in Molecular Biology (Clifton, N.J.)

|November 19, 2011

Summary

This summary is machine-generated.

Researchers improved ribosome display technology for protein evolution using the PURE system. This enhanced stability allows for reliable selection of functional peptides, such as those binding to antibodies.

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Published on: December 6, 2017

Area of Science:

Biotechnology
Molecular Biology
Protein Engineering

Background:

Ribosome display is a powerful technique for directed protein evolution and selection of functional polypeptides.
Traditional cell-free protein synthesis systems contain nucleases that degrade the messenger RNA-ribosome-polypeptide complex, limiting reliability.
Instability of the ternary complex hinders accurate selection of desired proteins.

Purpose of the Study:

To develop a more stable and controllable ribosome display system.
To overcome the limitations of nuclease activity in cell extract-based systems.
To enable reliable recovery and selection of functional polypeptides.

Main Methods:

Development of a modified ribosome display system utilizing the Protein Synthesizing Using Recombinant elements (PURE) system.
Leveraging the PURE system's low nuclease and inhibitory factor activity.
Selection of specific binding peptides from random libraries against monoclonal antibodies.

Main Results:

The PURE-based ribosome display system significantly enhances the stability of the messenger RNA-ribosome-polypeptide ternary complex.
Reliable recovery of selected mRNA was achieved due to the stabilized complex.
Successful selection of peptides with specific binding affinity to monoclonal antibodies was demonstrated.

Conclusions:

The modified PURE system offers a highly effective and controllable ribosome display platform.
This enhanced system overcomes previous limitations, enabling more reliable directed protein evolution.
The usability of the PURE system for ribosome display is strongly substantiated for protein selection applications.