Improvement of DNA/Metal Particle Adsorption in Tungsten-Based Biolistic Bombardment; Alkaline pH is Necessary for DNA Adsorption and Suppression of DNA Degradation

Yuya Yoshimitsu; Kiwamu Tanaka; Takashi Tagawa; Yasushi Nakamura; Tomoaki Matsuo; Shigehisa Okamoto

doi:10.1007/s12374-009-9068-0

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Improvement of DNA/Metal Particle Adsorption in Tungsten-Based Biolistic Bombardment; Alkaline pH is Necessary for DNA Adsorption and Suppression of DNA Degradation

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Improvement of DNA/Metal Particle Adsorption in Tungsten-Based Biolistic Bombardment; Alkaline pH is Necessary for DNA Adsorption and Suppression of DNA Degradation

Yuya Yoshimitsu, Kiwamu Tanaka, Takashi Tagawa, Yasushi Nakamura, Tomoaki Matsuo and Shigehisa Okamoto

Journal of plant biology = Singmul Hakhoe chi, Vol.52(6), pp.524-532

12/2009

DOI: https://doi.org/10.1007/s12374-009-9068-0

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https://hdl.handle.net/2376/104513

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Abstract

Life Sciences

Alkaline pH

DNA adsorption

Plant Systematics/Taxonomy/Biogeography

Plant Breeding/Biotechnology

Biolistic bombardment

Tungsten particle

DNA degradation

Plant Sciences

Plant Ecology

Plant Genetics & Genomics

Tungsten particles have long been used as microcarriers in biolistic bombardment because of their cost-effectiveness compared to alternative gold particles—even if the former have several drawbacks, including their DNA-degrading activity. We characterized tungsten-induced DNA degradation to assess the value of this metal particle and to improve tungsten-based biolistic bombardment. Alkaline pH, low temperature, and high salt concentration were found to diminish tungsten-induced DNA breakdown. The pH was the most influential factor in this phenomenon, both in aqueous solutions and on the particles. Furthermore, alkaline pH greater than 9.4 of an adsorption mixture was found to be essential for DNA binding to metal particles. Based on these findings, we propose a new formula of DNA/tungsten adsorption by using TE buffers that keep alkaline pH (>9.4) of the mixture, in which tungsten-bound plasmid DNA cleavage was suppressed to half the level of that in the conventional DNA-binding condition.

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Title: Improvement of DNA/Metal Particle Adsorption in Tungsten-Based Biolistic Bombardment; Alkaline pH is Necessary for DNA Adsorption and Suppression of DNA Degradation
Creators: Yuya Yoshimitsu - Department of Agricultural Sciences and Natural Resources Kagoshima University Kohrimoto 1-21-24 Kagoshima 890-0065 Japan
Kiwamu Tanaka - Department of Agricultural Sciences and Natural Resources Kagoshima University Kohrimoto 1-21-24 Kagoshima 890-0065 Japan
Takashi Tagawa - Department of Biochemical Science and Technology Kagoshima University Kohrimoto 1-21-24 Kagoshima 890-0065 Japan
Yasushi Nakamura - Department of Food Sciences and Nutritional Health Kyoto Prefectural University Shimogamo, Sakyo-ku Kyoto 606-8522 Japan
Tomoaki Matsuo - Department of Biochemical Science and Technology Kagoshima University Kohrimoto 1-21-24 Kagoshima 890-0065 Japan
Shigehisa Okamoto - Department of Agricultural Sciences and Natural Resources Kagoshima University Kohrimoto 1-21-24 Kagoshima 890-0065 Japan
Publication Details: Journal of plant biology = Singmul Hakhoe chi, Vol.52(6), pp.524-532
Academic Unit: Plant Pathology, Department of
Publisher: Springer-Verlag; New York
Identifiers: 99900546891601842
Language: English
Resource Type: Journal article