水稻谷蛋白基因GluC启动子的克隆及表达分析

doi:10.7525/j.issn.1673-5102.2018.06.016

摘要/Abstract

摘要： 水稻谷蛋白仅在水稻种子胚乳中表达，其启动子是分离胚乳特异性表达启动子的理想材料。本研究克隆了GluC基因启动子pGluC，生物信息学分析表明pGluC内部含有胚乳特异性表达所需要的Skn-1 motif和ACGT-box元件。将pGluC启动子和7个5'端缺失启动子片段构建到pGPTV-GUS载体上，转化水稻愈伤组织，进行组织化学染色和GUS酶活分析。结果表明：全长及截短的-1 911、-1 611、-1 311 bp启动子均能驱动GUS基因在水稻种子胚乳中高效稳定表达。-999、-451、-203、-102 bp启动子失去了胚乳表达特异性，在根、茎或者叶中也检测到GUS表达。该结果为实现外源目的基因在水稻胚乳中特异高效表达提供了理论依据。

关键词: 水稻, 启动子, 顺式作用元件, GUS组织化学染色, GUS酶活分析

Abstract: Rice glutelin, which is expressed in endosperm exclusively, is ideal candidate for isolating promoters with endosperm specificity. We isolated the GluC 5'-flanking sequence pGluC. Analysis of pGluC sequence by PlantCARE and PLACE revealed it contained a series of endosperm specific elements, such as Skn-1 motif and ACGT-box. Thus, we constructed the pGluC and truncated pGluC into pGPTV-GUS vector and transferred the vectors into rice. We performed histochemical assay and GUS activity detection. The full-length and -1 911,-1 611, -1 311 bp promoters were mainly accumulated in endosperm. The -999, -451, -203, -102 bp promoters were mainly accumulated in endosperm, and less strongly in roots, stems and leaves, which changed their endosperm-specific expression pattern. Our findings would contribute to the selection of a suitable promoter for crop molecular improvement.

Key words: rice, promoter, cis-acting element, GUS histochemical staining, quantitative GUS assay

中图分类号:

李文静, 孙艳香. 水稻谷蛋白基因GluC启动子的克隆及表达分析[J]. 植物研究, 2018, 38(6): 921-930.

LI Wen-Jing, SUN Yan-Xiang. Cloning and Expression Analysis of Glutelin GluC Gene Promoter in Rice[J]. Bulletin of Botanical Research, 2018, 38(6): 921-930.

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