Abstract
Electroporation was used for the delivery and subsequent expression of GUS and anthocyanin reporter genes into intact maize immature embryos. The optimal conditions consisted of culturing immature embryos for 4 days on N6 1-100-25-Ag medium prior to electroporation (375 V/cm; 960 µF capacitance) in EPR buffer containing DNA and 0.07 M sodium glutamate at room temperature (22°C) after a 10 min heat shock at 37°C. Under these conditions, over 40 spots of GUS transient activity were observed per immature embryo. Transient gene expression after electroporation was further demonstrated using an anthocyanin construct, which is specific for expression in plant cells.
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Abbreviations
- EPR:
-
electroporation buffer
- GUS:
-
β-glucuronidase
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
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Songstad, D.D., Halaka, F.G., DeBoer, D.L. et al. Transient expression of GUS and anthocyanin constructs in intact maize immature embryos following electroporation. Plant Cell Tiss Organ Cult 33, 195–201 (1993). https://doi.org/10.1007/BF01983234
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DOI: https://doi.org/10.1007/BF01983234