利用蛋白质组学技术鉴定大豆籽粒贮藏蛋白A1aB1b亚基缺失体  

Identification A_(1a)B_(1b)-subunit Deficiency of Glycinin in Soybean (Glycine max L.Merrill) Seed by Proteomics Approach

作　　者：刘春[1,2] 麻浩[1] 高文瑞[1] 王显生[1] 陈晨[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,国家大豆改良中心,江苏南京210095 [2]衡阳师范学院生命科学系,湖南衡阳421008

出　　处：《大豆科学》2010年第5期738-741,共4页Soybean Science

基　　金：引进国际先进农业计划资助项目(M2001-207);衡阳师范学院科学基金资助项目(09A40);湖南省高等学校科学研究资助项目(10C0493)

摘　　要：利用SDS-PAGE对220份大豆种子贮藏蛋白亚基含量进行筛选,在获得大豆籽粒贮藏蛋白11S组分组I亚基变异种质的基础上,经双向电泳分辨大豆贮藏蛋白亚基正常品种(南农大黄豆)和亚基变异品种(桂阳紫金豆)成熟种子总蛋白的蛋白质组。用PDQuest软件分析双向电泳凝胶图谱,发现在大豆贮藏蛋白11S酸性亚基位置有等电点和分子量分别为5.40和37.85kDa、5.24和37.2kDa、5.15和37.05kDa的蛋白质点在正常种子中表达而在变异种质种子中未表达。对这3个蛋白质点用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)测定其胶内酶解后的肽质量指纹谱(PMF),对获得的PMF用Mascot软件在NCBInr数据库中查询比对,鉴定出这3个蛋白质点均为大豆球蛋白A1aB1b亚基同源三聚体,表明变异种质种子缺少贮藏蛋白A1aB1b亚基。A mutant line was successfully selected from 220 soybean cultivars by using SDS-PAGE, which characterized by lack of a certain subonit of glycinin group I, seed protein proteome of the mutant line ＇ Guiyang zijindou＇ and cultivar ＇ Nannongdahuangdou＇ were compared using PDQuest software（Bio-Rad） after two-dimensional gel electrophoresis （2-DE） , with the cuhivar ＇ Nannongdahuangdou＇ as control. The results showed that three protein spots with pI 5.15, Mr 37.85 kDa; pI 5.24, Mr 37.2 kDa; pl 5.40, Mr 37.05 kDa respectively were present in seed protein of the cultivar ＇ Nannongdahuangdou＇ , however, no protein spots were presented in the location of glycinin group I acid subunits of the mutant line. These three protein spots were identified by MALDI- TOF- MS, and peptide mass fingerprinting data were queried in NCBInr database by Mascot. All these proteins were identified as crystal structure of soybean proglycinin A1aB1b homotrimer. The results indicate that the mutant line lacking A1aB1b-subunits.

关 键 词：11S蛋白 A1aB1b亚基 SDS—PAGE 蛋白质组学 

分 类 号：S565.1[农业科学—作物学]