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Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance

热1 libingbing 添加于 2010-4-14 22:23 | 1852 次阅读 | 0 个评论

作者
Zhang X, Liu S, Takano T
摘要
Two cysteine proteinase inhibitors (cystatins) from Arabidopsis thaliana, designated AtCYSa and AtCYSb, were characterized. Recombinant GST-AtCYSa and GST-AtCYSb were expressed in Escherichia coli and purified. They inhibit the catalytic activity of papain, which is generally taken as evidence for cysteine proteinase inhibitor function. Northern blot analyses showed that the expressions of AtCYSa and AtCYSb gene in Arabidopsis cells and seedlings were strongly induced by multiple abiotic stresses from high salt, drought, oxidant, and cold. Interestingly, the promoter region of AtCYSa gene contains a dehydration-responsive element (DRE) and an abscisic acid (ABA)-responsive element (ABRE), which identifies it as a DREB1A and AREB target gene. Under normal conditions, AtCYSa was expressed in 35S: DREB1A and 35S: AREB1 plants at a higher level than in WT plants, while AtCYSa gene was expressed in 35S: DREB2A plants at the same level as in WT plants. Under stress conditions (salt, drought and cold), AtCYSa was expressed more in all three transgenic plants than in WT plants. Over-expression of AtCYSa and AtCYSb in transgenic yeast and Arabidopsis plants increased the resistance to high salt, drought, oxidative, and cold stresses. Taken together, these data raise the possibility of using AtCYSa and AtCYSb to genetically improve environmental stresses tolerance in plants.
详细资料
- 文献种类:期刊
- 期刊名称: Plant Molecular Biology
- 期刊缩写: Plant Mol Biol
- 期卷页: 2008年第68卷第1-2期 131-143页
- ISBN: 0167-4412
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附件
Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance

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【原创】本文从拟南芥中克隆得到了两个cystatins——AtCYSa，AtCYSb，通过与GST融合表达E.coli，分析蛋白活性，证实具有抑制cys蛋白酶的功能。通过northern blot分析干旱，盐，氧化及冷胁迫下的表达特性。对拟南芥和酵母进行转化，证实确实在逆境中起到作用，另外，分析启动子，发现还有ARE及ABRE元件，通过对超表达35S: DREB1A，35S: AREB1，35S: DREB2A拟南芥下这两个基因的表达分析，推测目的基因与这三个调控元件的关系，最后还推测可能促进多种参与逆境的功能性蛋白及调节性蛋白表达提高抗性。总体来说，是一篇很好的论文，特别是讨论部分，写得很好，而且作者也在文后指出了不足，如作为cys蛋白酶抑制剂，应该分析超表达后cys蛋白酶活性的变化等。

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