来源于一粒系小麦具有表达活性的高分子量谷蛋白基因Ay的鉴定.pdfVIP

来源于一粒系小麦具有表达活性的 高分子量谷蛋白基因Ay 的鉴定与克隆 农学院 农学（农业产业化） 王峰 指导教师：魏育明 研究员 摘要：采用 SDS方法，对 20 份一粒系小麦（Einkorn wheat ，AA ，2n=2x=14 ）材料 高分子量谷蛋白亚基组成进行检测发现，PI277123 具有两条高分子量谷蛋白亚基。利用一 对特异引物对其基因编码区进行 PCR 扩增，同时扩增出两条 DNA 片段，大小分别是2.4kb u 和 1.8kb。将 1.8bp 片段克隆于 T-载体后获得阳性克隆 AY , 对基因完整编码区序列进行了 测定。根据核苷酸序列推导的氨基酸序列表明，AYu 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：一粒系小麦；高分子量麦谷蛋白亚基；分子克隆 Identification and Molecular Cloning of Ay-type HMW-GS Expressional Genes from Einkorn Wheat Student: Wang Feng (Agronomy College, Agronomy) Supervisor: Prof. Wei Yu Ming Abstract: HMW glutenin subunits in 20 Einkorn wheat (AA, 2n=2x=14) accessions were detected by SDS. In accession PI277123, there were two HMW glutenin subunits including an active y-type subunit. Two fragments (2.4kb and 1.8kb) corresponding to open reading frames (ORFs) of HMW glutenin genes were amplified with a pair of primers. After the amplified fragment (1.8kb) was cloned into the plasmid vector u u PMD18-T, a clone AY with corresponding insert sizes was obtained and sequenced. It was found that AY had u the typical primary structure of y-type subunit. The deduced amino acid sequence of AY has a signal peptide, consisting of the first 21 amino acids, followed by a non-repetitive N-terminal of 104 residues at 5’ terminal and a non-repetitive C-terminal domain with 42 residues at 3’ terminal. Five and o