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Plant and Cell Physiology Advance Access published online on October 30, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp154
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Identification of Zinc-Responsive Proteins in the Roots of Arabidopsis thaliana using a Highly Improved Method of Two-Dimensional Electrophoresis

Yoichiro Fukao1,*, Ali Ferjani3, Masayuki Fujiwara1, Yuka Nishimori1 and Iwao Ohtsu2

1 Plant Science Education Unit, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630-0101 Japan
2 Laboratory of Cell Biotechnology, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, 630-0101 Japan
3 Department of Biology, Tokyo Gakugei University, Tokyo, 184-8501, Japan

* To whom correspondence should be addressed Corresponding Author Yoichiro Fukao phone: +81-743-72-6250 fax: +81-743-72-6251 E-mail: fukao{at}bs.nasit.jp


  Abstract

Zinc (Zn) is an essential micronutrient for various physiological and metabolic processes in plants, although it is toxic in excess. To better understand Zn-responsive proteins, we developed a highly improved method of isoelectric focusing (IEF) in which whole lysate from Arabidopsis roots is subjected to IEF without any desalting steps. In this method, samples extracted with lysis buffer containing 1.5% SDS can also be directly applied to IEF. By applying this method to Zn-treated Arabidopsis roots, 10 upregulated and 17 downregulated proteins were identified, 15 of which showed a significant correlation with previously reported transcriptomic data.

Keywords: Arabidopsis thaliana - 2-D gel electrophoresis - IEF - microsomal fraction - SDS - Zn-stress

(Received September 11, 2009; Accepted October 21, 2009)
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