Biotechnology Bulletin ›› 2020, Vol. 36 ›› Issue (12): 247-255.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0555

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Sample Preparation and Data Analysis Method for Soybean Seed Proteome

MU Yong-ying(), WANG Dao-ping, CHEN Ming, QIU Li-juan, PAN Ying-hong()   

  1. Institute of Crop Science,Chinese Academy of Agricultural Sciences,Beijing 100081
  • Received:2020-05-09 Online:2020-12-26 Published:2020-12-22
  • Contact: PAN Ying-hong E-mail:mu_yongying@163.com;panyinghong@caas.cn

Abstract:

This study constructs a method for effective sample preparation and analysis of soybean seed proteomes via liquid mass spectrometry,aiming to improve the coverage of soybean proteome analysis. The mature seeds of soybean variety Zhonghuang 35 were used as materials to compare the protein extraction and enzyme digestion methods,liquid phase separation gradients and databases. The results showed that the identified proteins were the most when soybean seed proteome was extracted using modified urea thiourea buffer and digested with Lys-C/trypsin sequentially,then subjected on 90min nano liquid chromatographic separation and analyzed based on UniProt database. Finally,this method was applied to the preparation and analysis of mature seeds of soybean variety Tokachi-Nagaha,and a total of 2 244 non-redundant proteins were identified from three independent samples,in which about 61% could be repeatedly identified. Thus this method is reliable and stabile,and it is suitable for the proteome sample preparation of soybean seed and data analysis.

Key words: proteome, sample preparation, protein digestion, soybean seed