旱麦草属物种高分子量麦谷蛋白新基因的鉴定与分离.pdfVIP

旱麦草属物种高分子量麦谷蛋白新基因的鉴定与分离 农学院 生物技术专业 陈超 指导教师：郑有良 教授 魏育明 副研究员 摘 要 采用 SDS方法，对光稃旱麦草（Eremopyrum bonaepartis ，FF ， 2n=2x=14 ）材料PI261220 高分子量谷蛋白亚基组成进行检测发现，PI261220 具有两 条高分子量谷蛋白亚基。利用一对特异引物对其蛋白基因编码区进行PCR 扩增，同 时扩增出两条DNA 片段，大小分别是3.0 和 1.5kb。将两片段别克隆于T-载体后获 得阳性克隆pF3.0 和pF1.5, 对两端序列进行了测定。根据核苷酸序列推导的氨基酸 序列表明，Fy1.5 具有典型的y 型高分子量谷蛋白亚基基因特征，在5’端具有一个 由21 个氨基酸组成的信号肽和 104 个氨基酸组成的N 末端，在3’端有一个42 个 氨基酸组成的C 末端且在其N 和 C 末端分别有5 个和1 个半胱氨酸残基。 关键词 稃旱麦草（Eremopyrum bonaepartis ）；分子量麦谷蛋白亚基；分子克隆 Characterization of the high molecular weight glutenin subunit genes from Eremopyrum bonaepartis Abstract HMW glutenin subunits from Eremopyrum bonaepartis (FF, 2n=2x=14) accession PI261220 were detected by SDS. In accession PI261220, there were two HMW glutenin subunits, which exhibited unusual higher or lower electrophoretic mobility respectively. After the two amplified fragments (3.0 and 1.5kb) were cloned into the plasmid vector pMD18-T, two clones pFx3.0 and pFy1.5 with corresponding insert sizes were obtained. The results of partial DNA sequencing of two terminals showed that Fy1.5 had the typical primary structure of y-type subunit. Fy1.5 has a signal peptide, consisting of the first 21 amino acids, following a non-repetitive N-terminal of 104 residues at 5’ terminal and a non-repetitive C-terminal domain with 42 residues 3’ terminal. Five and one cysteine residues were observed in the N- and C-terminal, respectively. Key words Eremopyrum bonaepartis ；HWM glutenin subunit ；Molecular cloning 人类食用的70% 的蛋白质来源于粮食作物的种子，而世界粮食总产量的30%是小 [1] 麦 ，所以小麦种子蛋白质直接关系到人类的营养状况。成熟小麦种子的贮藏蛋白主 要包括麦谷蛋白（Glutenin ）和醇溶蛋白（Gliadin ）两大类。麦谷蛋白可分为高分子 量谷蛋白（High-molecular-weight glutenin subunit ，HWM-GS ）和低分子量麦谷蛋白 （Low-molecular-weight glutenin subunit ，LWM-GS ）。醇溶蛋白包括α﹑β﹑γ和ω 四 种组分。 高分子量麦谷蛋白亚基是由位于小麦第 1 部分同源群染色体长臂的 3 个基因位 点所控制，分别称位Glu-A1﹑Glu-B1 和 Glu-D