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Updated: May 15, 2026

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing

Published on: August 7, 2021

Defining the bacteroides ribosomal binding site.

Udo Wegmann¹, Nikki Horn, Simon R Carding

¹Gut Health and Food Safety Programme, Institute of Food Research, Norwich Research Park, Norwich, United Kingdom. udo.wegmann@ifr.ac.uk

Applied and Environmental Microbiology

|January 22, 2013

Summary

This summary is machine-generated.

Researchers developed new genetic tools for studying Bacteroides bacteria, which are abundant in the human gut. These tools enable better understanding of Bacteroides

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

The human gastrointestinal tract harbors numerous commensal microorganisms, with Bacteroides species being highly abundant in the colon.
Bacteroidetes possess unique transcription initiation signals, necessitating specialized genetic tools for their study.
Existing genetic tools are suboptimal for investigating the roles and functions of Bacteroides in the gut environment.

Purpose of the Study:

To develop novel genetic tools for enhanced protein expression in the genus Bacteroides.
To identify and characterize translation initiation signals specific to Bacteroides.
To facilitate research on the functions of Bacteroides in the human gastrointestinal tract.

Main Methods:

Construction of a series of expression vectors tailored for Bacteroides.
Evaluation of pepI from Lactobacillus delbrueckii subsp. lactis as a novel reporter gene.
Identification of the 3' end of Bacteroides ovatus 16S rRNA and analysis of its ribosomal binding site.

Main Results:

Development of expression vectors enabling varied protein expression levels in Bacteroides.
Successful use of pepI as a reporter gene in Bacteroides.
Characterization of a core ribosomal binding site sequence crucial for efficient translation in Bacteroides ovatus, incorporated into vector design.

Conclusions:

The developed genetic tools and characterized regulatory elements significantly improve the study of Bacteroides.
The findings on translation initiation signals are applicable to the broader order Bacteroidales.
These advancements will aid in elucidating the roles of these important gut bacteria.