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Mol. Cells 2009; 27(4): 423-428

Published online April 13, 2009

https://doi.org/10.1007/s10059-009-0060-9

© The Korean Society for Molecular and Cellular Biology

Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

Xu Xiaojing, Zhou Yijun, Wei Shanjun, Ren Dongtao, Yang Min, Bu Huahu, Kang Mingming, Wang Junli, and Feng Jinchao

Received: December 18, 2009; Revised: February 22, 2009; Accepted: February 26, 2009

Abstract

Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

Keywords expression, RACE, superoxide dismutase (SOD), ThCSD gene, Thellungiella halophila

Article

Research Article

Mol. Cells 2009; 27(4): 423-428

Published online April 30, 2009 https://doi.org/10.1007/s10059-009-0060-9

Copyright © The Korean Society for Molecular and Cellular Biology.

Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

Xu Xiaojing, Zhou Yijun, Wei Shanjun, Ren Dongtao, Yang Min, Bu Huahu, Kang Mingming, Wang Junli, and Feng Jinchao

Received: December 18, 2009; Revised: February 22, 2009; Accepted: February 26, 2009

Abstract

Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

Keywords: expression, RACE, superoxide dismutase (SOD), ThCSD gene, Thellungiella halophila

Mol. Cells
May 31, 2023 Vol.46 No.5, pp. 259~328
COVER PICTURE
The alpha-helices in the lamin filaments are depicted as coils, with different subdomains distinguished by various colors. Coil 1a is represented by magenta, coil 1b by yellow, L2 by green, coil 2a by white, coil 2b by brown, stutter by cyan, coil 2c by dark blue, and the lamin Ig-like domain by grey. In the background, cells are displayed, with the cytosol depicted in green and the nucleus in blue (Ahn et al., pp. 309-318).

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