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Updated: Jun 3, 2026
10:12
A Robotic Platform for High-throughput Protoplast Isolation and Transformation
Published on: September 27, 2016
The chloroplast transformation toolbox: selectable markers and marker removal
Anil Day1, Michel Goldschmidt-Clermont
1Faculty of Life Sciences, The University of Manchester, Manchester, UK.
Plant Biotechnology Journal
|March 24, 2011
Summary
Plastid transformation enables genetic engineering across many plant species using selectable markers. Efficient methods for removing these antibiotic resistance markers are crucial for biotechnological applications.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Genetics
Background:
- Plastid transformation is a key tool in plant biotechnology and basic research, applicable to diverse species beyond initial models like Chlamydomonas and tobacco.
- Efficient selection of fully transformed, polyploid plastid genomes is essential for successful transgenic line development.
- Various selection strategies exist, including photoautotrophy, antibiotic resistance, and herbicide tolerance, each with specific applications and limitations.
Purpose of the Study:
- To review and highlight the importance of efficient selectable markers in plastid transformation.
- To discuss the necessity and methods for removing selectable markers post-transformation.
- To present advanced techniques for marker gene removal in plastid engineering.
Main Methods:
- Review of established plastid transformation selection methods, including photoautotrophy and antibiotic resistance markers (e.g., aadA).
- Exploration of marker removal strategies utilizing homologous recombination, direct repeats, and transient co-integration/co-transformation.
- Investigation of site-specific recombinase systems for efficient marker gene excision from plastids.
Main Results:
- Antibiotic resistance markers, such as aadA, are widely used but often undesirable in final transgenic products.
- Homologous recombination-based methods and transient expression systems facilitate marker segregation and removal.
- Foreign site-specific recombinases offer an effective alternative for precise marker gene removal.
Conclusions:
- Efficient selectable markers are critical for advancing plastid transformation technology.
- Marker removal techniques are vital for optimizing plastid engineering for various applications.
- The development of sophisticated marker removal systems enhances the utility and applicability of plastid transformation.
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