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：7

作者：

Yoshimitsu, Yuya ^{[1
]}

Tanaka, Kiwamu ^{[1
]}

Tagawa, Takashi ^{[3
]}

Nakamura, Yasushi ^{[2
]}

Matsuo, Tomoaki ^{[3
]}

Okamoto, Shigehisa ^{[1
]}

机构：

[1] Kagoshima Univ, Dept Agr Sci & Nat Resources, Kagoshima 8900065, Japan

[2] Kyoto Prefectural Univ, Dept Food Sci & Nutr Hlth, Sakyo Ku, Kyoto 6068522, Japan

[3] Kagoshima Univ, Dept Biochem Sci & Technol, Kagoshima 890065, Japan

来源：

JOURNAL OF PLANT BIOLOGY | 2009年 / 52卷 / 06期

关键词：

Biolistic bombardment; Tungsten particle; Alkaline pH; DNA degradation; DNA adsorption; TRANSFORMATION; CELLS;

D O I：

10.1007/s12374-009-9068-0

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

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.

引用

页码：524 / 532

页数：9

共 19 条

[1] Efficient microprojectile bombardment-mediated transformation of rice using gene cassettes [J].

Breitler, JC ;

Labeyrie, A ;

Meynard, D ;

Legavre, T ;

Guiderdoni, E .

THEORETICAL AND APPLIED GENETICS, 2002, 104 (04) :709-719

[2]

Buchowicz J, 2003, MOLEC METH PLAN ANAL, V23, P175

[3]

Carmody W.R., 1961, Journal of Chemical Education, V38, P559, DOI DOI 10.1021/ED038P559

[4] Engineered GFP as a vital reporter in plants [J].

Chiu, WL ;

Niwa, Y ;

Zeng, W ;

Hirano, T ;

Kobayashi, H ;

Sheen, J .

CURRENT BIOLOGY, 1996, 6 (03) :325-330

[5]

GRENOBLE DC, 1983, HYDROCARBON CONVERSI

[6]

Jefferson RA., 1987, PLANT MOL BIOL REP, V5, P387, DOI [10.1007/BF02667740, DOI 10.1007/BF02667740]

[7] AN IMPROVED ASSAY FOR BETA-GLUCURONIDASE IN TRANSFORMED-CELLS - METHANOL ALMOST COMPLETELY SUPPRESSES A PUTATIVE ENDOGENOUS BETA-GLUCURONIDASE ACTIVITY [J].

KOSUGI, S ;

OHASHI, Y ;

NAKAJIMA, K ;

ARAI, Y .

PLANT SCIENCE, 1990, 70 (01) :133-140

[8] Relaxation, linearization and fragmentation of supercoiled circular DNA by tungsten microprojectiles [J].

Krysiak, C ;

Mazus, B ;

Buchowicz, J .

TRANSGENIC RESEARCH, 1999, 8 (04) :303-306

[9] Generation of DNA double-strand breaks and inhibition of somatic embryogenesis by tungsten microparticles in wheat [J].

Krysiak, C ;

Mazus, B ;

Buchowicz, J .

PLANT CELL TISSUE AND ORGAN CULTURE, 1999, 58 (03) :163-170

[10] Tungsten particle-induced nicking of supercoiled plasmid DNA [J].

Mazus, B ;

Krysiak, C ;

Buchowicz, J .

PLASMID, 2000, 44 (01) :89-93

← 1 2 →