DNA transformation into rice protoplasts and the gene expression in the regenerated plants

Research Project

Project/Area Number	62480029
Research Category	Grant-in-Aid for General Scientific Research (B)
Allocation Type	Single-year Grants
Research Field	Breeding science
Research Institution	TOHOKU UNIVERSITY
Principal Investigator	HINATA Kokichi Professor, Lab.Plt.Breed., Fac. of Agr., Tohoku Univ., 農学部, 教授 (00005589)
Co-Investigator(Kenkyū-buntansha)	TORIYAMA Kinya Assistance, Lab.Plt.Breed., Fac. of Agr., Tohoku Univ., 農学部, 助手 (20183882)
Project Period (FY)	1987 – 1988
Project Status	Completed (Fiscal Year 1988)
Budget Amount *help	¥6,400,000 (Direct Cost: ¥6,400,000) Fiscal Year 1988: ¥1,800,000 (Direct Cost: ¥1,800,000) Fiscal Year 1987: ¥4,600,000 (Direct Cost: ¥4,600,000)
Keywords	Rice / Oryza sativa L. / protoplast / electroporation / transgenic plant / APH(3')II gene / GUS gene / APH(3')II遺伝子 / トランジエント遺伝子発現
Research Abstract	Whole plants of rice (Oryza sativa L.) were derived from protoplasts, which had been electroporated with plasmid DNA possessing a chimeric gene encoding aminoglycoside phosphotransferase II (APH(3')II, conferring resistance to kanamycin and G418) and the cauliflower mosaic virus 35S promoter. Effective conditions of electroporation were determined by monitoring transient expression of the GUS gene (Clontech Lab. Inc.). Transformed calli were selected on the basis of tolerance to the antibiotic, G418. Several regenerated plants possessed functional APH(3')II activity due to the integration of intact foreign DNA into their genome as confirmed by Southern blot analysis.