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Updated: Jul 29, 2025

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants

Published on: July 1, 2018

Split selectable marker systems utilizing inteins facilitate gene stacking in plants.

Guoliang Yuan^1,2,3, Haiwei Lu^1,4, Kuntal De¹

¹Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37831, USA.

Communications Biology

|May 26, 2023

Summary

This summary is machine-generated.

Researchers developed split selectable marker systems using inteins for efficient plant gene stacking. This method allows for the simultaneous insertion of multiple genes into various plant species, overcoming limitations in current crop development techniques.

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Transient Gene Expression in Tobacco using Gibson Assembly and the Gene Gun

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Published on: April 18, 2014

Area of Science:

Plant biotechnology
Molecular biology
Genetics

Background:

Gene stacking is crucial for developing crops with improved traits.
Limited selectable marker options hinder efficient multi-gene insertion in plants.
Agrobacterium-mediated transformation is a key tool in plant genetic engineering.

Purpose of the Study:

To establish and validate split selectable marker systems utilizing inteins for plant co-transformation.
To demonstrate the efficacy of these systems in reconstituting visible markers and conferring antibiotic resistance.
To assess the general applicability of the split marker systems in model plants like Arabidopsis and poplar.

Main Methods:

Development of split selectable marker systems based on protein splicing inteins.
Agrobacterium-mediated co-transformation of plant tissues (tobacco, Arabidopsis, poplar).
Reconstitution of the visible RUBY marker from non-functional fragments.
Selection of transformed plants using split Kanamycin and Hygromycin resistance markers.

Main Results:

Successful reconstitution of the RUBY visible marker in tobacco via split marker system.
Demonstrated utility in Arabidopsis and poplar for stacking reporter genes (eYGFPuv, RUBY).
Effective use of split Kanamycin and Hygromycin resistance markers for plant selection.
Robust co-transformation achieved, enabling simultaneous insertion of multiple genes.

Conclusions:

Split selectable marker systems using inteins provide a robust method for plant co-transformation.
This technology efficiently enables the simultaneous insertion of multiple genes in both herbaceous and woody plants.
The developed systems offer a valuable tool to overcome limitations in current plant breeding and genetic engineering.