酸浆种子蛋白提取方法的比较研究

李晓琳; 邵爱娟; 陈 敏; 黄璐琦

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酸浆种子蛋白提取方法的比较研究

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- 酸浆种子蛋白提取方法的比较研究
- Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis
- 中国实验方剂学杂志 2010年16卷第1期页码：14-17
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- 作者简介：
- 基金信息：
  
  国家科技基础条件平台项目(2005DKA21004)
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- 纸质出版日期：2010
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李晓琳, 邵爱娟, 陈敏, 等. 酸浆种子蛋白提取方法的比较研究[J]. 中国实验方剂学杂志, 2010,16(1):14-17.

LI Xiao-lin, SHAO Ai-juan, CHEN Min, et al. Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis[J]. Chinese journal of experimental traditional medical formulae, 2010, 16(1): 14-17.
李晓琳, 邵爱娟, 陈敏, 等. 酸浆种子蛋白提取方法的比较研究[J]. 中国实验方剂学杂志, 2010,16(1):14-17. DOI：

LI Xiao-lin, SHAO Ai-juan, CHEN Min, et al. Comparison of Protein Extraction Methods for the Semen Seu Fructus Physalis[J]. Chinese journal of experimental traditional medical formulae, 2010, 16(1): 14-17. DOI：

摘要

目的 :建立适用于酸浆种子的SDS-PAGE和2-D PAGE蛋白样品提取方法。方法: 比较直接提取法、TCA-丙酮法、Tris-丙酮法、饱和酚法、改良饱和酚法等7种提取方法所获酸浆种子总蛋白含量以及SDS-PAGE电泳的谱带数目、强度和电泳分辨率等方面的差异。结果: HEPES buffer直接提取法提取蛋白效率较高

杂质少

电泳谱带清晰

分辨率高;PVP buffer和Lysis buffer A直接提取法次之

但杂质较多;TCA-丙酮法、Tris-丙酮法、饱和酚法和改良的饱和酚法差别不大

蛋白纯度较高

但提取效率低

谱带数目少;此外

Lysis buffer A法、丙酮法和酚法易产生高丰度蛋白的干扰。结论: 直接提取法中的HEPES buffer法最好

优于丙酮法、酚法和其它直接提取法。

Abstract

Objective: To compare seven protein extraction protocols(PVP buffer

HEPES buffer

Lysis buffer A extraction and TCA/acetone

tris-base/acetone

phenol/ammonium acetate

acetone/phenol/ammonium acetate precipitation) and establish a suitable one for total proteins extraction from Semen Seu Fructus Physalis. Methods: The protein yield

protein purity and SDS-PAGE patterns was determined and analyzed. Results: HEPES buffer extraction produce the purer sample

much more clear bands and highest resolution. PVP buffer and Lysis buffer A extraction had most impurities. TCA/acetone

tris-base/acetone

phenol/ammonium acetate

acetone/phenol/ ammonium acetate precipitation methods had the purest sample

but protein yield and band numbers were lowest. Besides

the methods employing Lysis buffer was easy to produce high-abundance proteins and interfered in the electrophoresis resolution. Conclusion: HEPES buffer extraction was the best sample preparation protocol for SDS-PAGE and 2-D PAGE.

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酸浆种子蛋白提取方法的比较研究