Sample Preparation and Data Analysis Method for Soybean Seed Proteome

doi:10.13560/j.cnki.biotech.bull.1985.2020-0555

Abstract

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

MU Yong-ying, WANG Dao-ping, CHEN Ming, QIU Li-juan, PAN Ying-hong. Sample Preparation and Data Analysis Method for Soybean Seed Proteome[J]. Biotechnology Bulletin, 2020, 36(12): 247-255.

Figures/Tables 7

References 32

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液相梯度	蛋白	肽段	二级谱图数	肽段-谱图匹配数
30 min	709±11 b	2 890±53 c	13 015±129 c	4 747±124 c
60 min	1 364 b±4 a	5 620±22 b	36 751±173 b	10 966±30 b
120 min	1 396±47 a	5 771±60 a	53 129±301 a	25 256±694 a

液相梯度	蛋白	肽段	二级谱图数	肽段-谱图匹配数
30 min	709±11 b	2 890±53 c	13 015±129 c	4 747±124 c
60 min	1 364 b±4 a	5 620±22 b	36 751±173 b	10 966±30 b
120 min	1 396±47 a	5 771±60 a	53 129±301 a	25 256±694 a

数据库名称	数据库总蛋白条目数	鉴定蛋白数
数据库名称	数据库总蛋白条目数	有功能注释	无功能注释
Phytozome：Gmax_275_Wm82.a2.v1	88 647	1 411	560
UniProt：Glycine max_cv.Williams 82	88 687	1 237	753
NCBI：Glycine_max_v2.1	88 647	1 501	457

数据库名称	数据库总蛋白条目数	鉴定蛋白数
数据库名称	数据库总蛋白条目数	有功能注释	无功能注释
Phytozome：Gmax_275_Wm82.a2.v1	88 647	1 411	560
UniProt：Glycine max_cv.Williams 82	88 687	1 237	753
NCBI：Glycine_max_v2.1	88 647	1 501	457