孟 敏,强琴琴,高 翔,陈其皎,董 剑.小麦Waxy蛋白亚基毛细管电泳分离体系的构建[J].麦类作物学报,2015,35(12):1653 |
小麦Waxy蛋白亚基毛细管电泳分离体系的构建 |
Separation System of Waxy Protein Subunits in Wheat by Capillary Electrophoresis |
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DOI:10.7606/j.issn.1009-1041.2015.12.07 |
中文关键词: 小麦 Waxy蛋白 毛细管电泳 缓冲液 |
英文关键词:Wheat Waxy protein Capillary electrophoresis Running buffer |
基金项目:国家现代农业产业技术体系专项(CARS-3-2-47);国家自然科学基金项目(30900896);中央高校基本科研业务费专项(QN2009007);西北农林科技大学唐仲英育种基金项目(A212020912) |
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中文摘要: |
为构建Waxy蛋白亚基的快速、高效、高通量毛细管电泳分离鉴定体系,为该亚基理化特性的分析、功能的发掘及含Waxy蛋白亚基种质的快速筛选提供参考依据,以小麦中国春为材料,采用逐一优化毛细管电泳体系参数的方法,系统研究了毛细管电泳中缓冲液各成分以及pH值对中国春中Waxy蛋白亚基出峰效果的影响,初步构建了普通小麦Waxy蛋白亚基的最佳分离体系。结果表明,在检测波长为214 nm、以0.05 mol·L-1硼砂+8% 乙腈+0.8% SDS 溶液(pH 9.5)为缓冲液、分离电压20 kV、温度25 ℃、进样压力0.5 psi×5 s时,在6~9 min处出现峰高约0.015 au的三个主峰,且图形清晰,基线水平,重复性好。本研究构建的毛细管电泳系统分离体系可有效分离普通小麦Waxy蛋白亚基。 |
英文摘要: |
In order to lay a foundation for follow up research of wheat Waxy protein subunits, effects of pH and buffer components of capillary electrophoresis on peaks of Waxy protein subunits in Chinese Spring were studied systematically. And an efficient capillary electrophoresis separation system of Waxy protein subunits of hexaploid wheat was intended to constructed. Results showed that three peaks with 0.015 au height, clear graph, horizontal baseline and excellent repeatability appeared at 214 nm detection wavelength, 20 kV separation voltage, 25 ℃ and 0.5 psi×5 s sampling pressure with 0.05 mmol·L-1 borax + 8% acetonitrile + 0.8% SDS solution buffer (pH 9.5). The results implied that an efficient capillary electrophoresis separation system of Waxy protein subunits of hexaploid wheat was constructed successfully. |
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