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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
被引:0
|作者:
Yuya Yoshimitsu
Kiwamu Tanaka
Takashi Tagawa
Yasushi Nakamura
Tomoaki Matsuo
Shigehisa Okamoto
机构:
[1] Kagoshima University,Department of Agricultural Sciences and Natural Resources
[2] Kyoto Prefectural University,Department of Food Sciences and Nutritional Health
[3] Kagoshima University,Department of Biochemical Science and Technology
来源:
Journal of Plant Biology
|
2009年
/
52卷
关键词:
Biolistic bombardment;
Tungsten particle;
Alkaline pH;
DNA degradation;
DNA adsorption;
D O I:
暂无
中图分类号:
学科分类号:
摘要:
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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页码:524 / 532
页数:8
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