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克隆和功能分析玉米中一个应激诱导基因(Zmap)的启动子。

Cloning and functional analysis of the promoter of a stress-inducible gene (Zmap) in maize.

机构信息

College of Veterinary Medicine, Northeast Agricultural University, Harbin, P. R. China.

Heilongjiang Key Laboratory for Animal Disease Control and Pharmaceutical Development, Harbin, P. R. China.

出版信息

PLoS One. 2019 Feb 8;14(2):e0211941. doi: 10.1371/journal.pone.0211941. eCollection 2019.

Abstract

The anionic peroxidases play an important role in a variety of plant physiological processes. We characterized and isolated the Zmap promoter (PZmap) at the 5' flanking region in order to better understand the regulatory mechanisms of Zmap gene expression. A series of PZmap deletion derivatives, termed a1 -a6, at positions -1694, -1394, -1138, -784, -527 and -221 from the translation start site were blended to the β-glucuronidase reporter gene. Agrobacterium-mediated transformation method was used to study each deletion construct in tobaccos. Sequence analysis showed that several cis-acting elements (MYB binding site, Box-II, a TGACG-element, a CGTCA-element and a low temperature responsive element) were located within the promoter. Deletion analysis suggested the sequence between -1,694 and -1394bp may contain cis-elements associated with GUS up regulation. The MYB binding site (-757) might act as a negative drought-responsive element. There might be repressor elements located in the region (-1,694 to -1394bp) to repress Zmap expression under 4°C. The characterized promoter would be an ideal candidate for genetic engineering for improving the resistance of maize to different stressors.

摘要

阴离子过氧化物酶在多种植物生理过程中发挥着重要作用。我们对 Zmap 启动子(PZmap)在 5'侧翼区域进行了特征分析和分离,以便更好地理解 Zmap 基因表达的调控机制。在翻译起始位点处的-1694、-1394、-1138、-784、-527 和-221 位置,一系列的 PZmap 缺失衍生片段,称为 a1 -a6,被融合到β-葡萄糖醛酸酶报告基因中。我们使用农杆菌介导的转化方法在烟草中研究了每个缺失构建体。序列分析表明,启动子内存在几个顺式作用元件(MYB 结合位点、Box-II、TGACG 元件、CGTCA 元件和低温响应元件)。缺失分析表明,-1694 至-1394bp 之间的序列可能包含与 GUS 上调相关的顺式元件。MYB 结合位点(-757)可能作为一个负的干旱响应元件发挥作用。在 4°C 下,可能存在位于-1694 到-1394bp 区域的抑制子元件来抑制 Zmap 的表达。该特征化的启动子将是用于遗传工程以提高玉米对不同胁迫抗性的理想候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/918d/6368321/0de6314cf24b/pone.0211941.g001.jpg

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