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番茄早期光诱导蛋白基因强启动子的功能表征。

Functional characterization of a strong promoter of the early light-inducible protein gene from tomato.

机构信息

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, 142290, Russia.

Nikita Botanical Gardens-National Scientific Center, Russian Academy of Sciences, Yalta, 298648, Russia.

出版信息

Planta. 2019 Oct;250(4):1307-1323. doi: 10.1007/s00425-019-03227-x. Epub 2019 Jul 3.

Abstract

The tomato ELIP gene promoter is mainly active in the ripening fruit. Considering its high activity, the promoter could be used for molecular breeding of plants in the future. The ability to obtain new varieties of transgenic plants with economically valuable traits relies on a high level of target gene expression, which is largely controlled by a gene promoter. Hence, research aimed at finding and characterizing new tissue-specific promoters that direct gene expression in specific plant tissues or at certain developmental stages has become the most important field of plant biotechnology. Here, we cloned and characterized the promoter of the early light-inducible protein (ELIP) gene from tomato (Solanum lycopersicum cv. Yalf). ELIPs are produced in the presence of light and putatively function in the chloroplast-to-chromoplast conversion, playing a photorepairing role in the photosynthetic system. Analysis of the promoter sequence revealed multiple cis-acting elements related to light responsiveness, and other motifs involved in plant hormone response and circadian control. To determine the functionality of the promoter, seven 5'-deletion variants were fused with the β-glucuronidase (GUS) reporter gene and introduced into tomato. Histochemical analysis of transgenic tomato plants revealed different levels of GUS activity in most analyzed tissues, depending on the promoter fragment used. The intensity of staining was considerably higher in ripening fruits than in unripe and non-fruit tissues. Quantitative analysis indicated that the level of GUS activity with the longest (full-length) version of the ELIP promoter in ripened fruits was comparable to that in plants expressing the constitutive CaMV35S promoter. Further, the location of both negative and positive regulatory motifs was identified. The described ELIP promoter is a potential tool for various applications in plant biotechnology.

摘要

番茄 ELIP 基因启动子主要在成熟果实中活跃。考虑到其高活性,该启动子将来可用于植物的分子育种。获得具有经济价值性状的转基因植物新品种的能力依赖于目标基因表达的高水平,而这在很大程度上受基因启动子控制。因此,寻找和表征新的组织特异性启动子,以指导基因在特定植物组织或特定发育阶段表达,已成为植物生物技术最重要的领域。在这里,我们从番茄(Solanum lycopersicum cv. Yalf)中克隆和表征了早期光诱导蛋白(ELIP)基因的启动子。ELIPs 在光照下产生,推测其在叶绿体-叶绿体转化中发挥作用,在光合作用系统中起光修复作用。启动子序列分析揭示了多个与光反应相关的顺式作用元件,以及其他与植物激素反应和昼夜节律控制有关的基序。为了确定启动子的功能,将七个 5'-缺失变体与β-葡萄糖醛酸酶(GUS)报告基因融合,并导入番茄中。对转基因番茄植株的组织化学分析表明,在所分析的大多数组织中,根据使用的启动子片段,GUS 活性水平不同。与未成熟和非果实组织相比,成熟果实中的染色强度要高得多。定量分析表明,在成熟果实中使用最长(全长)版本的 ELIP 启动子时,GUS 活性的水平与表达组成型 CaMV35S 启动子的植物相当。此外,还确定了负调节和正调节基序的位置。所描述的 ELIP 启动子是植物生物技术中各种应用的潜在工具。

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