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Related Experiment Video

Updated: May 9, 2026

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Agrobacterium-Mediated Immature Embryo Transformation of Recalcitrant Maize Inbred Lines Using Morphogenic Genes

Published on: February 14, 2020

Gene tagging with engineered Ds elements in maize.

Yubin Li¹, Gregorio Segal, Qinghua Wang

¹Waksman Institute, Rutgers University, Piscataway, NJ, USA.

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

|August 7, 2013

Summary

Researchers developed a new method for maize gene isolation using Dissociation (Ds) transposons and green fluorescent protein (GFP) markers. This technique facilitates gene discovery by enabling PCR amplification of DNA adjacent to tagged genes.

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Last Updated: May 9, 2026

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Published on: February 27, 2021

Area of Science:

Plant molecular biology
Genetics and genomics
Maize (Zea mays) research

Background:

Efficient gene isolation is crucial for understanding plant genetics and for crop improvement.
Traditional gene tagging methods can be labor-intensive and may lack precision.
Dissociation (Ds) transposons are mobile genetic elements used for gene tagging in maize.

Purpose of the Study:

To describe protocols for isolating maize genes utilizing Dissociation (Ds) transposons.
To leverage a green fluorescent protein (GFP) transgene as a marker for Ds elements.
To establish a genetic resource for targeted gene tagging across the maize genome.

Main Methods:

Introduction of Ds transposons engineered with a green fluorescent protein (GFP) transgene into maize.
Phenotypic scoring of Ds element activity using GFP fluorescence.
Polymerase Chain Reaction (PCR) amplification of DNA sequences adjacent to Ds insertion sites using unique primers.

Main Results:

The GFP marker allows for easy identification and phenotypic scoring of Ds transposition events.
Unique primers anchored to the Ds element facilitate the isolation of flanking genomic DNA via PCR.
A strategy is being implemented to mobilize Ds elements from various genomic locations, creating a comprehensive resource.

Conclusions:

The described protocols offer an efficient method for maize gene isolation and characterization.
The GFP-marked Ds transposon system provides a powerful tool for genetic analysis and gene discovery in maize.
The developed genetic resource aims to enable targeted gene tagging throughout the maize genome, aiding functional genomics studies.