Abstract
Several methods of transformation are currently available for delivering exogenous DNA into animal and plant cells. In this study, a novel and efficient transformation system for DNA delivery/expression with a capacity to transport DNA of high molecular weight was developed. This system can overcome the shortcomings of traditional transformation methods such as Agrobacterium-mediated transformation, particle bombardment, and the electroporation method. The method developed in this study uses calcium alginate micro beads to immobilize DNA molecules in combination with polyethylene glycol treatment. In addition, it is simple and low-cost, and requires limited equipment. Using this method, we have successfully transformed tobacco plants, screening by kanamycin resistance. The transformed genes in the transformants were confirmed by PCR and Southern hybridization.
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Acknowledgements
We would like to thank Dr. Yasuo Niwa (Shizuoka University) and Dr. Mitsuyasa Hasebe (NIBB) for kindly providing us with the pUC18-sGFP and pTN80 plasmids, respectively. This study was supported by Basic Research Activities for Innovative Biosciences, Japan.
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Liu, H., Kawabe, A., Matsunaga, S. et al. Obtaining transgenic plants using the bio-active beads method. J Plant Res 117, 95–99 (2004). https://doi.org/10.1007/s10265-003-0141-3
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DOI: https://doi.org/10.1007/s10265-003-0141-3