Related Experiment Video

Updated: Dec 3, 2025

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays

Published on: November 12, 2012

RedEx: a method for seamless DNA insertion and deletion in large multimodular polyketide synthase gene clusters

Chaoyi Song¹, Ji Luan¹, Ruijuan Li¹

¹State Key Laboratory of Microbial Technology, Institute of Microbial Technology, Helmholtz International Lab for Anti-infectives, Shandong University-Helmholtz Institute of Biotechnology, Shandong University, Qingdao, Shandong, 266237, China.

Nucleic Acids Research

|October 29, 2020

Summary

A new RedEx method enables seamless DNA manipulation in large polyketide synthase gene clusters. This technique facilitates the diversification of chemical structures, like modifying the spinosad insecticide, for various applications.

Area of Science:

Synthetic biology
Molecular biology
Biochemistry

Background:

Polyketide synthase (PKS) genes contain large repetitive DNA sequences, complicating seamless manipulation.
Diversifying chemical structures through biosynthesis reprogramming is crucial for discovering novel compounds.

Purpose of the Study:

To develop a method for seamless DNA manipulation in large PKS gene clusters.
To demonstrate the utility of this method for modifying the spinosad gene cluster.

Main Methods:

Developed the RedEx method, combining Redαβ-mediated homologous recombination, ccdB counterselection, and exonuclease-mediated in vitro annealing.
Applied RedEx to refactor the 79-kb spinosad gene cluster to replace an ethyl group with a butenyl group at C-21.
Utilized RedEx to seamlessly delete the rhamnose 3'-O-methyltransferase gene.

Main Results:

Successfully inserted an exogenous extension module into the PKS gene without extra sequences.
Generated spinosad derivatives with a butenyl group at C-21.
Produced rhamnosyl-3'-desmethyl spinosad derivatives by gene deletion.

Conclusions:

The RedEx method provides a powerful tool for seamless mutagenesis in large and complex DNA sequences.
This technique facilitates the rational design of PKS gene clusters for chemical structure diversification.
RedEx offers significant advantages for genetic engineering and synthetic biology applications.

More Related Videos

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028

Published on: January 13, 2017

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

Exon Recombination

Exon Recombination

The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon...

Articles linked to this work by shared authors, journal, and citation graph.

Orchestrating triacylglycerol biosynthesis and hydrolysis via synthetic genetic circuits enhances spinosad production in Streptomyces albus.

Bioresource technology·2026

Development of an efficient genome editing system using recombinase-mediated cassette exchange in E. coli Nissle 1917 for large gene cluster integration and heterologous astaxanthin production.

Synthetic and systems biotechnology·2026

Construction and modification of a low-copy plasmid-based infectious clone for GI-19 genotype IBV via Red/ET recombineering: A simplified and efficient reverse genetics system for coronavirus.

Poultry science·2026

Heterologous production of epothilones in Streptomyces albus J1074 via promoter engineering of the biosynthetic gene cluster and precursor supply enhancement.

Synthetic and systems biotechnology·2026

Virulence attenuation of intestinal pathogenicity via combined gene deletion in duck enteritis vaccine strain restores gut microbiota balance and enhances safety.

Poultry science·2026

Safety and immunogenicity of a broad-spectrum HVT vector vaccine for avian infectious bursal disease.

Poultry science·2025

See all related articles