Direct Gene Transfer into Mature Seeds via Electroporation
Direct Gene Transfer into Mature Seeds via Electroporation
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Direct Gene Transfer into Mature Seeds via Electroporation
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– Direct gene transfer after vacuum treatment.
– For research use, patent royalty is free in Japan.
– Authors are ready for patent licensing negotiation.
Basic Method
1. Soak mature seeds in water overnight.
2. Transfer the seeds into electroporation buffer.
3. Vacuum treatment.
4. Electroporation on ice.
5. Incubate the seeds on ice for a few hours.
6. Grow the plants under the favorable conditions.
7. Select the transformed plants using antibiotics.
Fig. 1. Electroporation Apparatus and the Chamber (Produced by NEPA GENE Co., Ltd.)
Apparatus: CUY21EDIT/NEPA21
Chamber: CUY495P10
Electroporation on ice
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.
1.Starting materials are mature seeds.
2.Transformed plants are obtained in relatively short time.
3.Plant tissue culture is not necessary.
4.Possibly applicable to animal cells and microorganisms.
Fig. 2. Production of Transgenic Wheat 
GUS Gene ExpressionSelection by GeneticinTransformed Plants
Fig. 3. Transient GUS Gene Expression
Rice (japonica)…………………………. Rice (indica) ………….. Brassica
……….Soybean
Fig. 4.Gene transfer into Silk Worm Eggs
GFP Expression Fluorescence
Plant Genetic Engineering Unit, National Institute of Agrobiological Sciences; Takashi Hagio
*Improvement research is in progress in his laboratory.
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