摘 要 在SDS-PAGE筛选到A1aB1b或A1bB2或A2B1a亚基缺失的基础上,运用双向聚丙烯酰胺凝胶电泳(2-DE)分析大豆贮藏亚基正常品种南农大黄豆和亚基缺失品种桂阳紫金豆种子总蛋白的蛋白质组。差异蛋白质组学显示,等电点和分子量分别约为x1和y1、x2和y2、x3和y3的蛋白质点在正常品种种子中表达而在缺失品种中未表达。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(Matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)测定其胶内酶解后的肽质量指纹谱(Peptide Mass Fingerprint,PMF),经Mascot:Peptide Mass Fingerprint中NCBI数据库查询后,鉴定这3个蛋白质均为Soybean Proglycinin A1ab1b Homotrimer,表明桂阳紫金豆种子贮藏蛋白缺少A1ab1b亚基。
关键词 蛋白质组学; 鉴定; 大豆贮藏蛋白; A1aB1b亚基; 缺失
Abstract:This thesis focuses on applying 2-DE to analyze the proteome of the seed protein of soybean varieties “Nannongdahuangdou” which contains normal subunits and “Guiyangzijindou” which lacks subunits on the basis of SDS-PAGE selection and the absence of the A1aB1b or A1bB2 or A2B1a –subunits. Three protein spots with pIx1 , Mr y1kDa; pIx2 , Mr y2kDa; pIx3, Mr y3 kDa respectively show normal expression in the seed protein of the variety “Nannongdahuangdou”, while no expression in the mutant line. After being tested the PMF with Matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS and surveyed by the NCBI state base of Mascot:Peptide Mass Fingerprint, these three proteins were all identified as Soybean Proglycinin A1ab1b Homotrimer. The results indicated that the variety “Guiyangzijindou” lacks A1ab1b-sununits.
Keywords: Proteomics; identification; soybean storage protein; A1ab1b-sununits; absence