[Plant Seed] Direct Gene Transfer into Mature Seeds via Electroporation
[Plant Seed] Direct Gene Transfer into Mature Seeds via Electroporation
[Plant Seed] Direct Gene Transfer into Mature Seeds via Electroporation
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Direct gene transfer after the vacuum treatment
For research use, patent royalty is free in Japan
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Basic Method |
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Soak mature seeds in water overnight |
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Transfer the seeds into the electroporation buffer |
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Vacuum treatment |
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Electroporation on ice |
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Incubate the seeds on ice for a few hours |
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Grow the plants under the relevant favourable conditions |
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Select the transformed plants using antibiotics. |
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Fig. 1. Electroporation Apparatus and the Chamber (Produced by NEPA GENE Co., Ltd.)
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| Apparatus: NEPA21/CUY21EDIT |
Chamber: CUY495P10 |
Electroporation on ice |
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| Fertile transgenic plants were regenerated and self-fertilized seeds were obtained in rice and wheat. Transgene integration was confirmed by Southern hybridization. Transmission of the transgene into the next generation (T1) was indicated by PCR analysis. Transient gene expression was observed in several plant species and silk worm eggs. |
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Starting materials are mature seeds |
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Transformed plants are obtained in relatively short time |
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Plant tissue culture is not necessary |
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Possibly applicable to animal cells and microorganisms |
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Fig. 2. Production of Transgenic Whea
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| GUS Gene Expression |
Selection by Geneticin |
Transformed Plants |
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Fig. 3. Transient GUS Gene Expression
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| Rice (japonica) |
Rice (indica) |
Brassica |
Soybean |
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Fig. 4. Gene transfer into Silk Worm Eggs
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Takashi Hagio, Plant Genetic Engineering Unit, National Institute of Agrobiological Sciences
*Improvement research is in progress in his laboratory |
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