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Updated: Jun 22, 2026

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
Published on: March 28, 2018
Developmentally regulated site-specific marker gene excision in transgenic B. napus plants
Lilya Kopertekh1, Inge Broer, Joachim Schiemann
1Julius Kühn Institute, Federal Research Centre for Cultivated Plants (JKI), Institute for Biosafety of Genetically Modified Plants, Erwin-Baur-Str. 27, 06484, Quedlinburg, Germany.
Researchers created a novel Cre-vector system for Brassica napus, enabling the removal of marker genes. This method successfully generated marker-free transgenic plants, offering a valuable tool for crop improvement.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Genetics
Background:
- Marker genes are essential for identifying transgenic organisms but can pose challenges in subsequent breeding.
- Efficient methods for marker gene removal are crucial for developing commercially viable transgenic crops.
Purpose of the Study:
- To develop and validate a self-excision Cre-vector system for removing marker genes in Brassica napus.
- To assess the efficiency of marker gene excision based on transgene copy number.
Main Methods:
- Construction of a Cre-vector with lox-flanked marker genes and Cre recombinase.
- Agrobacterium-mediated transformation of Brassica napus.
- Segregation and molecular analysis (PCR, Southern blot) of transgenic progeny.
Main Results:
- Successful simultaneous excision of Cre recombinase and marker genes in Brassica napus.
- Detection of marker-free plants across all tested transgenic lines.
- Recombination efficiency ranged from 13% to 81%, influenced by transgene copy number.
Conclusions:
- The developed self-excision Cre-vector system is effective for generating marker-free transgenic Brassica napus.
- This technology holds potential for creating marker-free transgenic plants in sexually reproducing species.
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