Journal of Plant Biology

, Volume 52, Issue 6, pp 524–532

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

Authors

  • Yuya Yoshimitsu
    • Department of Agricultural Sciences and Natural ResourcesKagoshima University
  • Kiwamu Tanaka
    • Department of Agricultural Sciences and Natural ResourcesKagoshima University
  • Takashi Tagawa
    • Department of Biochemical Science and TechnologyKagoshima University
  • Yasushi Nakamura
    • Department of Food Sciences and Nutritional HealthKyoto Prefectural University
  • Tomoaki Matsuo
    • Department of Biochemical Science and TechnologyKagoshima University
    • Department of Agricultural Sciences and Natural ResourcesKagoshima University
Original Research

DOI: 10.1007/s12374-009-9068-0

Cite this article as:
Yoshimitsu, Y., Tanaka, K., Tagawa, T. et al. J. Plant Biol. (2009) 52: 524. doi:10.1007/s12374-009-9068-0

Abstract

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.

Keywords

Biolistic bombardment Tungsten particle Alkaline pH DNA degradation DNA adsorption

Abbreviations

cc

closed circular

GFP

green fluorescent protein

oc

open circular

Spd

spermidine-free base

TE

Tris–HCl/EDTA

Copyright information

© The Botanical Society of Korea 2009