普通小麦品种豫麦34和郑丰5号ω-醇溶蛋白基因的克隆与序列分析  

Cloning and Sequence Analysis ofω-gliadin Genes from Common Wheat(Triticum aestivum L.)Cultivars Yumai 34 and Zhengfeng 5

作　　者：李黎[1] 刘凌云[1] 李锁平[1] 李玉阁[1] 

机构地区：[1]河南大学生命科学学院,河南开封475004

出　　处：《麦类作物学报》2014年第9期1178-1184,共7页Journal of Triticeae Crops

基　　金："十二五"农村领域国家科技计划项目(2011BAD07B00;2012AA101105);国家自然科学基金面上项目(31271713)

摘　　要：ω-醇溶蛋白是影响小麦加工品质和诱发乳糜泻（celiac disease,CD）的面筋蛋白之一。为给小麦加工品质的分子改良和预防乳糜泻提供参考依据,利用3对特异引物,采用基因组PCR法,从优质小麦品种豫麦34和郑丰5号中克隆获得41个ω-醇溶蛋白序列。NCBI BLAST分析表明,克隆序列与已知序列的相似度均在91%-99%之间,推断其为ω-醇溶蛋白基因家族的基因。开放阅读框识别分析表明,11个序列（Y34W-1-Y34W-7,ZFW-1-ZFW-4）具有完整的开放阅读框,可编码203-377个氨基酸残基。ZFW-4和Y34W-1在重复区的插入片段中存在1个额外的半胱氨酸残基。CD免疫肽识别分析表明,11个基因中均分布有3种ω-醇溶蛋白的T细胞免疫肽,且肽段PQQPFPQQ存在多个拷贝。聚类分析显示,ω-醇溶蛋白具有一定的基因组特异性,11个克隆的基因与尾状山羊草（Aegilops markgrafii）和粗山羊草（Aegilops tauchii）的ω-醇溶蛋白基因序列具有相对较高的同源性。The ω -gliadins are one of the gulten proteins, which not only are correlated to wheat process quality but also are the active proteins in triggering celiac disease （CD）. In order to provide reference for wheat quality improvement and CD prevention, 41 unique clones （designated as Y34W 1-Y34W 23 and ZFW 1-ZFW 18, with GenBank accessions of KC716065-KC716082 and KF412579-KF412601） in total, were obtained from two common wheat cultivars Yumai34 and Zhengfeng 5 using a PCR based strategy. NCBI blast showed that the 41 cloned sequences had a highly homologous rate of 91%-99% with the genes in GenBank and they were inferred as the members of ω gliadins gene family. Open reading frame analysis demonstrated that 11 （designated as Y34W 1-Y34W 7, ZFW 1-ZFW 4） of the 41 sequences were novel full ORF ω gliadin genes which could encode polypeptides with 203 377 amino acid residues. Comparative analysis on the deduced amino acids sequences showed that the 11 full ORF genes had the typical structure of ω gliadins except that 2 genes （ZFW 4 and Y34W 1） possessed an additional cysteine residue in the repetitive domain. CD toxicity analysis showed that all the three known immunogenic epitopes, especially 2 5 peptides of PQQPFPQQ were presented in the 11 proteins. Homological analysis on the deduced amino acid sequences among the 11 novel genes and other 32 genes in public database showed that obvious differences occurred according to their origin of genome,and the 11 genes cloned in this work had relative highly homology to the ω gliadins derived from Aegilops markgrafii and Aegilops tauchii.

关 键 词：普通小麦 ω-醇溶蛋白 基因克隆 序列分析 

分 类 号：S512.1[农业科学—作物学] S330