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作者	单位
刘宝龙，姜燕燕，曹东，常砚梓，李云	(1.中国科学院西北高原生物研究所/青海省作物分子育种重点实验室，青海西宁 810000； 2.青海大学农林科学院，青海西宁 810000）

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中文摘要:

种子特异性启动子的克隆和功能分析不仅有助于阐明作物种子发育和胚乳特异性基因的表达调控机制，也是进行作物品质改良和种子生物反应器在内的植物基因工程改造的基础。本研究从大麦品种Golden pormise中克隆到大麦胚乳特异性启动子HorD，生物信息学分析表明，其具备启动子的基本元件，如A-box、TATA-box、CAAT-box等，此外还含有胚乳特异性表达所需要的元件Prolamin-box。为验证HorD启动子的表达特性，以pU1300载体为骨架，将HorD启动子和新型冠状病毒（SARA-CoV-2）刺突蛋白基因LPS通过双酶切连接的方式构建表达载体phorD-LPS，并利用农杆菌介导的遗传转化试验验证其种子表达特性。结果表明，HorD启动子能驱动LPS基因在转基因大麦各组织中表达，但在根、茎、叶及颖壳中表达量较低，在种子中表达量较高。这说明HorD启动子可以驱动外源目的基因在大麦种子中特异高效表达。

英文摘要:

The cloning and functional analyses of seed-specific promoters not only help to clarify the mechanisms of crop seed development and expression regulation of endosperm-specific genes, but also provide the basis for plant genetic engineering, including crop quality improvement and crop seed reactors. In this study, the barley endosperm-specific promoter HorD was cloned from the barley cultivar Golden promise. Bioinformatics analysis showed that the promoter not only had the basic elements such as A-box TATA-box, and CAAT-box, but also contained the element Prolamin-box, which was endosperm-specific expressed element. In order to verify the expression characteristics of HorD promoter, the vector phorD-LPS was constructed with restriction enzyme digestion with HorD promoter and the cloned novel coronavirus(SARA-CoV-2) spike protein gene LPS through double enzyme ligand used pU1300 vector as the skeleton.Its seed expression characteristics were verified by Agrobacterium-mediated stable genetic transformation experiments. The results showed that HorD promoter can drive the expression of LPS gene in transgenic barley tissues, and the expression level was lower in roots, stems, leaves and glume, but higher in seeds. The results above indicated that the HorD promoter can drive the specific and efficient expression of foreign gene of interest in barley seeds.

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