Obtaining transgenic plants using the bio-active beads method.
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.
Liu
Haibo
H
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Kawabe
Akira
A
Matsunaga
Sachihiro
S
Murakawa
Tomoko
T
Mizukami
Atsushi
A
Yanagisawa
Masanobu
M
Nagamori
Eiji
E
Harashima
Satoshi
S
Kobayashi
Akio
A
Fukui
Kiichi
K
D000464:Alginates / Q000494:pharmacology; D003470:Culture Media; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D007103:Immobilization; D007612:Kanamycin / Q000494:pharmacology; D007613:Kanamycin Resistance; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D010957:Plasmids; D011092:Polyethylene Glycols / Q000494:pharmacology; D011523:Protoplasts; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic
D000464:Alginates; D003470:Culture Media; D006603:Hexuronic Acids; D010940:Plant Proteins; D011092:Polyethylene Glycols; D007612:Kanamycin; D020723:Glucuronic Acid