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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (20): 30-34.

所属专题： 生物技术； 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

奶牛血浆蛋白质组学研究中双向凝胶电泳条件的优化

牛丽莉,魏彩虹,李宏滨,杜立新   

  • 收稿日期:2009-03-23 修回日期:2009-04-20 出版日期:2009-10-20 发布日期:2009-10-20
  • 基金资助:

    国家自然科学基金

Optimization on two-dimensional electrophoresis for plasma proteomic study in dairy cow

  • Received:2009-03-23 Revised:2009-04-20 Online:2009-10-20 Published:2009-10-20

PDF (PC)

978

摘要：

摘要：【研究目的】血浆蛋白组成复杂、信息含量丰富。血浆蛋白质结构和数量的改变，对疾病诊断和治疗具有重要的意义；【方法】双向电泳技术是蛋白质组研究中比较常用的一种蛋白质分离技术。根据血浆蛋白动态范围大，高丰度蛋白对分析存在干扰的特点，设计了两种实验方案，并比较了这两种方案的优缺点，；另外对双向电泳过程中的血浆样品的采集，样品制备，等电聚焦，聚丙烯酰胺电泳和图像分析步骤的注意事项进行讨论，为血浆作为诊断蛋白质组的应用提供参考。【结果】使用热SDS法处理样品可分离出230个可以重复的蛋白点。Aurum Affi-Gel Blue Mini columns除白蛋白柱处理样品分离了156个可重复的蛋白点；【结论】热SDS法处理血浆样品联合pH4-7的IPG胶条的方案，步骤简单，分离效果好。

关键词: 小白菜, 小白菜, NO-3-N, NH+4-N, 磷, 钾, 硝酸盐

Abstract:

Abstract： 【OBJECTIVE】The component of plasma is complex and plasma protein present information-rich. The change of structure and quantity of plasma protein is very important for disease diagnosis and treatment; 【METHOD】Two-dimensional electrophoresis is a key protein separation technology in proteomics research. Plasma protein have high dynamic range characterization, and high-abundance proteins can interfere with analysis. So we designed two experiments for study of plasma proteomics in dairy cow, and compared them in order to refer to plasma application as diagnosis marker;【RESULTS】230 protein points were separated which appeared in three repeat-gels by Hot-SDS method and 156 protein points were separated by Aurum Affi-Gel Blue Mini columns method.【CONCLUSION】The method that Hot-SDS and pH4-7 IPG strip were used was better.