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Updated: Jan 19, 2026

Optogenetic Control of Gene Expression in Corynebacterium glutamicum

Optogenetic Control of Gene Expression in Corynebacterium glutamicum

Published on: October 29, 2025

Multiple large segment deletion method for Corynebacterium glutamicum.

Nobuaki Suzuki¹, Hiroshi Nonaka, Yota Tsuge

¹Microbiology Research Group, Research Institute of Innovative Technology for the Earth (RITE), Kizugawadai, Kizu-Cho, Soraku-Gun, Kyoto, Japan.

Applied Microbiology and Biotechnology

|April 22, 2005

Summary

This summary is machine-generated.

Scientists developed a precise genome engineering method using Cre/loxP and double-strand breaks (DSBs) for scarless gene deletion. This technique successfully removed large genomic segments in Corynebacterium glutamicum without affecting growth.

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Genome engineering is crucial for understanding gene function and metabolic pathway optimization.
Precise and efficient methods for large-scale genomic modifications are needed.

Purpose of the Study:

To develop a scarless genome excision method for precise, large-segment deletions.
To demonstrate the feasibility of multiple, successive deletions in the Corynebacterium glutamicum genome.

Main Methods:

Utilized the Cre/loxP system combined with double-strand break (DSB)-stimulated intramolecular recombination.
Employed the I-SceI restriction endonuclease to induce DSBs for targeted recombination.
Performed sequential deletions of 14-, 43-, and 10-kb genomic regions.

Main Results:

Successfully achieved precise and scarless excision of large genomic regions (up to 67 kb).
Confirmed deletion accuracy through marker gene loss, PCR, and sequencing.
The resulting mutant, lacking 73 genes, maintained normal growth under standard conditions.

Conclusions:

The developed method enables efficient, scarless, and large-scale genome engineering.
This technique is valuable for manipulating bacterial genomes, particularly for strain improvement and synthetic biology applications.