文章摘要
植物水孔蛋白PIP2表达量的快速无标记检测
Label-free Detection of Expression Quantity of Plant PIP2 Protein
投稿时间:2019-03-15  修订日期:2019-05-09
DOI:
中文关键词: 水孔蛋白  PIP2  类肽适配体  表面等离激元共振成像  快速无标记检测
英文关键词: Aquaporin  PIP2  Peptoid  SPRi  Label-free detection method
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
作者单位E-mail
徐铭 大连理工大学 1084682834@mail.dlut.edu.cn 
赵子健   
夏秀英 大连理工大学 xx47@dlut.edu.cn 
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中文摘要:
      相较于传统的抗体检测,适配体更易于大量快速合成,且可和多种检测技术相结合,在蛋白检测方面具有巨大的潜力。水孔蛋白作为生物体内水分跨膜运输的主要途径,了解其表达量的变化在植物水代谢研究中有着重要意义。本研究利用混合列分法构建了8个残基的类肽适配体文库,结合表面等离激元共振成像技术,筛选得到能特异性结合高等植物水孔蛋白PIP2的类肽适配体PPA7(PIP2-binding peptoid aptamer 7),其亲和力KD高达2.52×10-9 mol/L。利用PPA7检测了石竹玻璃化试管苗和正常试管苗的水孔蛋白表达量,结果表明,玻璃化试管苗的水孔蛋白表达量显著高于正常试管苗。本研究提供了一种新的植物蛋白的定量检测策略,也为进一步明确水孔蛋白在组培苗玻璃化发生中的作用奠定了基础。
英文摘要:
      Compared with traditional antibody detection techniques, aptamers are easier to be rapidly synthesized in large quantities, and are easy to be combined with a variety of detection techniques, which has great potential in protein detection. As the main way of water transport across the membrane, it is of great significance to understand the changes of aquaporin expression in plant water metabolism. In this paper, an eight-mer peptoid library with one constant cysteine residue on the C-terminal was synthesized using the conventional split-and-pool method, and the peptoid aptamer PPA7 (PIP2-binding peptoid aptamer 7) with high affinity and specific binding to higher plant aquaporin (PIP2) was screened by surface plasmon resonance imaging (SPRi) technology. Its affinity was up to 2.52×10-9 mol/L. PPA7 was used to detect the expression of aquaporin in hyperhydric and normal plants. The results showed that the expression of aquaporin in hyperhydric plantlets was higher than that in normal ones. This paper provided a new quantitative detection strategy for plant proteins and laid a foundation for further clarifying the role of aquaporin in hyperhydricity.
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