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Related Experiment Video

Updated: May 14, 2026

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

Trackable multiplex recombineering for gene-trait mapping in E. coli.

Thomas J Mansell¹, Joseph R Warner, Ryan T Gill

¹Department of Chemical and Biological Engineering, Engineering Center, University of Colorado Boulder, Boulder, CO, USA.

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

|February 19, 2013

Summary

Trackable multiplex recombineering (TRMR) enables gene-trait mapping in Escherichia coli by creating thousands of simulated gene mutants. This method facilitates high-throughput screening for desired traits, advancing genome engineering and microbial research.

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Last Updated: May 14, 2026

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

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Published on: November 12, 2012

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

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Published on: January 8, 2015

Area of Science:

Microbiology
Molecular Biology
Genomics

Background:

Homologous recombination techniques have advanced genome engineering in Escherichia coli.
Multiplex recombineering offers powerful tools for modifying bacterial genomes.

Purpose of the Study:

To present trackable multiplex recombineering (TRMR), a novel method for gene-trait mapping in E. coli.
To enable the creation of large-scale mutant libraries for high-throughput screening.

Main Methods:

Combines oligonucleotide synthesis with multiplex recombineering to generate mutant libraries.
Utilizes DNA barcodes within recombineering cassettes for strain identification.
Employs high-throughput screening and selection for identifying strains with specific traits.

Main Results:

Developed two E. coli strain libraries totaling over 8,000 mutants.
TRMR allows for simulated gene knockdown and overexpression across the E. coli genome.
DNA microarray analysis enables rapid characterization of mutant populations.

Conclusions:

TRMR is an effective strategy for comprehensive gene-trait mapping in E. coli.
The method significantly enhances genome engineering capabilities for microbial research.
Facilitates discovery of gene functions through large-scale mutant analysis.