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马铃薯几丁质酶C基因的启动子将表达导向表皮细胞。

The promoter of the potato chitinase C gene directs expression to epidermal cells.

作者信息

Ancillo Gema, Hoegen Erika, Kombrink Erich

机构信息

Department of Plant Microbe Interactions, Max Planck Institute for Plant Breeding Research, Carl-von-Linné-Weg 10, 50829 Köln, Germany.

出版信息

Planta. 2003 Aug;217(4):566-76. doi: 10.1007/s00425-003-1029-0. Epub 2003 May 6.

Abstract

Chitinases are ubiquitous proteins that occur in all plants in multiple isoforms. We have isolated the ChtC2 gene encoding an unusual, basic (class I) chitinase from potato ( Solanum tuberosum L.). In contrast to other chitinase genes, ChtC2 is not activated by infection, but rather constitutively expressed in leaves and stems where it is restricted to epidermal cells. Sequence analysis revealed a number of potential regulatory elements in the promoter, but most striking was the presence of a 319-bp direct repeat located between -333 and -968 upstream of the transcription start site. For a functional analysis, a 1,322-bp promoter fragment and two 5' deletions of 782 bp and 162 bp in length were translationally fused to the beta-glucuronidase (GUS) reporter gene and used for transient expression studies by particle bombardment. All promoter constructs conferred expression of GUS activity in different epidermal cell types of potato leaves. Expression in parenchyma cells of the leaf mesophyll was not detectable with any of the ChtC2 gene promoter constructs, in contrast to the pattern observed with the 35S promoter from cauliflower mosaic virus. The epidermis-specific expression of the reporter gene was confirmed using transgenic potato plants containing the fusion of the entire ChtC2 promoter with the GUS reporter. Histochemical analysis indicated that the promoter was only active in epidermal cells of leaves.

摘要

几丁质酶是普遍存在的蛋白质,以多种同工型存在于所有植物中。我们从马铃薯(Solanum tuberosum L.)中分离出了编码一种不寻常的碱性(I类)几丁质酶的ChtC2基因。与其他几丁质酶基因不同,ChtC2不会被感染激活,而是在叶片和茎中组成型表达,且仅限于表皮细胞。序列分析揭示了启动子中一些潜在的调控元件,但最引人注目的是在转录起始位点上游-333至-968之间存在一个319 bp的直接重复序列。为了进行功能分析,将一个1322 bp的启动子片段以及两个长度分别为782 bp和162 bp的5'缺失片段与β-葡萄糖醛酸酶(GUS)报告基因进行翻译融合,并通过粒子轰击用于瞬时表达研究。所有启动子构建体均在马铃薯叶片的不同表皮细胞类型中赋予了GUS活性表达。与花椰菜花叶病毒35S启动子所观察到的模式相反,用任何ChtC2基因启动子构建体均未检测到叶肉薄壁细胞中的表达。使用含有完整ChtC2启动子与GUS报告基因融合体的转基因马铃薯植株证实了报告基因的表皮特异性表达。组织化学分析表明该启动子仅在叶片的表皮细胞中具有活性。

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