上海大学学报(自然科学版) ›› 2012, Vol. 18 ›› Issue (4): 430-434.doi: 10.3969/j.issn.1007-2861.2012.04.019

• 生命科学 • 上一篇    下一篇

苏丹草种子醇溶蛋白提取和电泳条件优化

邓志瑞,孙晶晶,张文举,陈沁   

  1. (上海大学 生命科学学院,上海 200444)
  • 出版日期:2012-08-30 发布日期:2012-08-30
  • 通讯作者: 邓志瑞(1964~),男,副教授,博士,研究方向为植物生理与分子生物学. E-mail:dengzhirui@staff.shu.edu.cn

Optimization of Extraction and Electrophoresis of Prolamin from Sorghum sudanense seeds

DENG Zhi-rui,SUN Jing-jing,ZHANG Wen-ju,CHEN Qin   

  1. (School of Life Sciences, Shanghai University, Shanghai 200444, China)
  • Online:2012-08-30 Published:2012-08-30

摘要: 以苏丹草(Sorghum sudanense) 种子为材料,以蛋白质提取率和电泳图谱效果为标准,分析不同提取剂、种子粉末质量(g)与提取剂体积(mL)的比例、还原剂二硫苏糖醇(dithiothreito,DTT)的浓度对种子醇溶蛋白提取效果的影响,同时研究上样量、不同胶浓度对SDS-PAGE检测蛋白的影响.结果表明,苏丹草的最佳提取剂为含有0.2%DTT的70%正丙醇,种子粉末质量与提取剂体积的最佳比例为1:8.在上样量适当的情况下,12%分离胶电泳可以得到清晰的电泳图谱.以苏丹草种子为材料所建立的醇溶蛋白提取和电泳条件,也可应用于其他几种常见的禾本科种子醇溶蛋白的分析,并可得到较好的分辨率.

关键词: SDS-PAGE, 醇溶蛋白提取, 苏丹草

Abstract: This paper studies the effects of different extraction solution, ratio of sampleweight to extractionsolution and concentrations of reduction reagent dithiothreito (DTT) on extraction efficiency and quality of electrophoresis of prolamin from seeds of Sorghum sudanense. Contribution of the gel concentration and loading dosage to the results of SDSPAGE  were analyzed by evaluating the electrophoresis. The results show that 70% propylalcohol supplemented with 0.2% DTT is the best extraction solution and the ratio of sampleweight to extractionsolution 1∶8 (g/mL) can give optimal results. SDSPAGE of 12% produces the best clearance for electrophoresis under a loading appropriate sample. Established condition based on prolamin from seeds of Sorghum sudanense can be applied to isolation and electrophoresis for prolamins from several other gramineae seeds with good results.

Key words: Sorghum sudanense, prolamin extraction, SDS-PAGE

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