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胡杨金属硫蛋白样 ThMT3,一种活性氧清除剂,增加了转基因酵母对 Cd(2+)、Zn(2+)、Cu(2+)和 NaCl 的耐受性。

Tamarix hispida metallothionein-like ThMT3, a reactive oxygen species scavenger, increases tolerance against Cd(2+), Zn(2+), Cu(2+), and NaCl in transgenic yeast.

机构信息

Key Laboratory of Forest Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, Harbin 150040, People's Republic of China.

出版信息

Mol Biol Rep. 2011 Mar;38(3):1567-74. doi: 10.1007/s11033-010-0265-1. Epub 2010 Sep 12.

Abstract

A metallothionein-like gene, ThMT3, encoding a type 3 metallothionein, was isolated from a Tamarix hispida leaf cDNA library. Expression analysis revealed that mRNA of ThMT3 was upregulated by high salinity as well as by heavy metal ions, and that ThMT3 was predominantly expressed in the leaf. Transgenic yeast (Saccharomyces cerevisiae) expressing ThMT3 showed increased tolerance to Cd(2+), Zn(2+), Cu(2+), and NaCl stress. Transgenic yeast also accumulated more Cd(2+), Zn(2+), and NaCl, but not Cu(2+). Analysis of the expression of four genes (GLR1, GTT2, GSH1, and YCF1) that aid in transporting heavy metal (Cd(2+)) from the cytoplasm to the vacuole demonstrated that none of these genes were induced under Cd(2+), Zn(2+), Cu(2+), and NaCl stress in ThMT3-transgenic yeast. H(2)O(2) levels in transgenic yeast under such stress conditions were less than half those in control yeast under the same conditions. Three antioxidant genes (SOD1, CAT1, and GPX1) were specifically expressed under Cd(2+), Zn(2+), Cu(2+), and NaCl stress in the transgenic yeast. Cd(2+), Zn(2+), and Cu(2+) increased the expression levels of SOD1, CAT1, and GPX1, respectively, whereas NaCl induced the expression of SOD1 and GPX1.

摘要

从柽柳叶片 cDNA 文库中分离到一个金属硫蛋白样基因 ThMT3,其编码一个 3 型金属硫蛋白。表达分析表明,ThMT3 的 mRNA 受高盐和重金属离子的上调诱导,并且主要在叶片中表达。表达 ThMT3 的转基因酵母(酿酒酵母)对 Cd(2+)、Zn(2+)、Cu(2+)和 NaCl 胁迫表现出更高的耐受性。转基因酵母还积累了更多的 Cd(2+)、Zn(2+)和 NaCl,但不积累 Cu(2+)。分析有助于将重金属(Cd(2+))从细胞质转运到液泡的四个基因(GLR1、GTT2、GSH1 和 YCF1)的表达表明,在 ThMT3 转基因酵母中,这些基因在 Cd(2+)、Zn(2+)、Cu(2+)和 NaCl 胁迫下均未被诱导。在相同条件下,这些胁迫条件下转基因酵母中的 H2O2 水平低于对照酵母中的水平的一半。三个抗氧化基因(SOD1、CAT1 和 GPX1)在 Cd(2+)、Zn(2+)、Cu(2+)和 NaCl 胁迫下在转基因酵母中特异性表达。Cd(2+)、Zn(2+)和 Cu(2+)分别增加了 SOD1、CAT1 和 GPX1 的表达水平,而 NaCl 诱导了 SOD1 和 GPX1 的表达。

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