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Cloning, Characterization and Expression Analysis of a Cytosolic Ascorbate Peroxidase Gene from Grimmia Pilifera

机译:格里姆氏菌胞质抗坏血酸过氧化物酶基因的克隆,鉴定及表达分析

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Ascorbate peroxidase(APX) is a class I peroxidase that catalyzed the conversion of H_2O_2 to H_2O and O_2 using ascorbate as the specific electron donor. This enzyme has a key function in scavenging reactive oxygen species (ROS) and the protection against toxic effects of ROS in plants. Here we reported the identification of an APX gene from a cDNA library of Grimmia pilifera. The full-length of the gene was cloned by 3'-RACE approach, and named as GpAPX. The predicted proteins of GpAPX consisted of 256 amino acid residues with a calculated molecular mass of 28.2 kDa, and displayed high similarity to other plant proteins. It also contains three conserved domains and two signatures. QRT-PCR analysis suggested that the expression of GpAPX gene was induced in both dehydration and rehydration. Its expression level increased under drought stress, inhibited at the begining of re-watering, and peaked on 12h after re-watering. The results demonstrated that the expression of GpAPX gene closely related to dehydration and rehydration, and its isolation established a good foundation for further study on the resistance of Grimmia pilifera to drought stress.
机译:抗坏血酸过氧化物酶(APX)是一类过氧化物酶,它以抗坏血酸盐为特定的电子供体,催化H_2O_2向H_2O和O_2的转化。该酶在清除活性氧(ROS)和防止ROS在植物中产生毒性作用方面具有关键作用。在这里,我们报道了从Grimmia pilifera的cDNA文库中鉴定出APX基因的过程。通过3'-RACE法克隆了该基因的全长,并命名为GpAPX。 GpAPX的预测蛋白由256个氨基酸残基组成,计算的分子量为28.2 kDa,与其他植物蛋白具有高度相似性。它还包含三个保守域和两个签名。 QRT-PCR分析表明,脱水和补液均诱导了GpAPX基因的表达。它的表达水平在干旱胁迫下增加,在再浇水开始时受到抑制,并在再浇水后12h达到峰值。结果表明,GpAPX基因的表达与脱水和补液密切相关,其分离为进一步研究Grimmia pilifera对干旱的抗性奠定了良好的基础。

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