Obtaining transgenic plants using the bio-active beads method.

Haibo Liu; Akira Kawabe; Sachihiro Matsunaga; Tomoko Murakawa; Atsushi Mizukami; Masanobu Yanagisawa; Eiji Nagamori; Satoshi Harashima; Akio Kobayashi; Kiichi Fukui

doi:10.1007/s10265-003-0141-3

PubMed · 15108034

Obtaining transgenic plants using the bio-active beads method.

Haibo Liu·Akira Kawabe·Sachihiro Matsunaga·Tomoko Murakawa·Atsushi Mizukami·Masanobu Yanagisawa·Eiji Nagamori·Satoshi Harashima·Akio Kobayashi·Kiichi Fukui

View original source ↗Export metadata ↓

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.

Explore related subjects

Alginates Culture Media Glucuronic Acid Hexuronic Acids Immobilization Kanamycin Kanamycin Resistance Plant Proteins Plants, Genetically Modified Plasmids Polyethylene Glycols Protoplasts Nicotiana Transformation, Genetic Journal Article Research Support, Non-U.S. Gov't

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Haibo Liu, Akira Kawabe, Sachihiro Matsunaga, Tomoko Murakawa, Atsushi Mizukami, Masanobu Yanagisawa, Eiji Nagamori, Satoshi Harashima, Akio Kobayashi, Kiichi Fukui. 2004-02-13. Obtaining transgenic plants using the bio-active beads method.. https://doi.org/10.1007/s10265-003-0141-3

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2007

Protein engineering of Acidovorax facilis 72W nitrilase for bioprocess development.

Hydroxycarboxylic acid monomers can be used to prepare industrially important polymers. Enzymatic production of such hydroxycarboxylic acids is often preferred to chemical production since the reactions are run at ambient temperature, do not require strongly acidic or basic reaction conditions, and produce the desired product with high selectivity at high conversion. However, native enzymes often do not perform desired reactions with the efficiency required for commercial applications. Protein engineering was used to significantly increase the specific activity of nitrilase from Acidovorax facilis 72W for the conversion of 3-hydroxyvaleronitrile to 3-hydroxyvaleric acid. Overexpression of engineered nitrilase enzymes in Escherichia coli, combined with immobilization of whole cells in alginate beads that can be recycled many times has facilitated the development of a commercially viable bioprocess for production of 3-hydroxyvaleric acid.

Alginates↗

PubMed2007

Zeta-potentials of alginate-PLL capsules: a predictive measure for biocompatibility?

Alginate-poly-L-lysine (PLL) microencapsulation of cells is a promising approach to prevent rejection in the absence of immunosuppression. Clinical application, however, is hampered by insufficient insight in factors influencing biocompatibility of the capsules. By now, it has been accepted that not only the chemical composition of the materials applied but also other factors contribute to bioincompatibility. The zeta-potential serves as a measure for the electrical charge of the surface and has been shown to be a predictive value for the interfacial reactions between the biomaterial and the surrounding tissue in other applications. In the present study, we have assessed the streaming potential of alginate-PLL capsules composed of either low-, intermediate-, or high-guluronic (G) alginate to calculate the zeta-potential. The zeta-potentials of the capsules were compared to the biological response against the capsules at 4 weeks after implantation in the rat. We show that high-G and low-G alginates provoke a more severe response in the rat than capsules prepared of intermediate-G alginate. This correlates with a higher zeta-potential of the high-G and low-G alginates and by a change in zeta-potential at lower pH. These lower pH-levels are common directly after implantation as the consequence of a host-response associated with mandatory surgery. Our results suggest that we should not only consider the capsule properties under physiological circumstances to explain bioincompatibility but also the capsule features during common pathophysiological situations.

Alginates↗

PubMed2007

Effect of immersion disinfection with Perform-ID on alginate, an alginate alternative, an addition-cured silicone and resultant type III gypsum casts.

OBJECTIVE: This study investigated the effect of a commonly used immersion disinfectant upon three different impression materials and any subsequent effects on the abrasion resistance, hardness and surface detail reproduction of gypsum casts. DESIGN: A laboratory study. MATERIALS AND METHODS: Under standardised conditions a total of 120 impressions were made of a ruled test block using irreversible hydrocolloid (Alginoplast), an 'alginate alternative' addition-cure silicone (Position Penta) and a conventional addition-cure silicone (President). The impressions were examined for surface detail reproduction prior to and after disinfection with Perform-ID. The type III casts were evaluated for surface detail reproduction, surface hardness and abrasion resistance. RESULTS: (1) None of the disinfected alginate specimens could reproduce the 50 microm line. (2) Casts produced from the disinfected alginate were significantly less hard than from disinfected Position Penta and President (P <0.001). (3) Disinfection significantly affected the abrasion resistance of casts made from Position Penta (P = 0.029). (4) Disinfection did not significantly affect President or its subsequent casts (P >0.05). CONCLUSION: If disinfecting with Perform-ID, the impression should be made with a conventional addition-cured silicone if good surface detail reproduction of the impression material and a hard and abrasion resistant type III gypsum cast are required.

Alginates↗