Cloning of nine γ-gliadin mRNAs (cDNAs) from wheat and the molecular characterization of comparative transcript levels of γ-gliadin subclasses
Cloning of nine γ-gliadin mRNAs (cDNAs) from wheat and the molecular characterization of comparative transcript levels of γ-gliadin subclasses
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DOI：
10.1016/j.jcs.2005.07.002
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发表时间：
2006-01-01
影响因子：
3.8
通讯作者：
Barro, F
中科院分区：
文献类型：
--
作者：
Pistón, F;Dorado, G;Barro, F
从面包和硬粒小麦的总mRNA中获得了9个新的伽马醇溶蛋白全长cDNA序列。开放阅读框(ORF)的长度范围为720~936bp，分别对应于239~311个氨基酸残基的推测蛋白质。根据系统发育数据重建的所有九个序列都显示了与GenBank注册的伽马醇溶蛋白相似的序列。所有已知的伽马-醇溶蛋白序列在由邻接法构建的树中分为四个主要组。底漆。设计了定量实时聚合酶链式反应(QRT-PCR)对面包小麦籽粒发育过程中这四个伽马醇溶蛋白组进行区分。两个面包小麦品系的表达谱显示，组II的伽马醇溶蛋白的基因转录水平较高，而组I的伽马醇溶蛋白的表达水平最低。因此，第二组的伽马醇溶蛋白在‘Perico’和‘Anza’中的表达分别比其他伽马醇溶蛋白高4倍和10倍。所报道的结果可能有助于理解发育中的面包小麦籽粒中γ-醇溶蛋白的合成和沉积顺序。此外，四组间表达水平的差异可用于选择具有特定y-醇溶蛋白谱的基因类型。这些发现也将有助于在转基因小麦中上调和下调伽马-醇溶蛋白基因的表达。(C)2005爱思唯尔有限公司。保留所有权利。
Nine new full-length cDNA sequences for gamma-gliadins were generated from total mRNA from bread and durum wheat. The length of the open reading frames (ORFs) ranged from 720 to 936 bp, which corresponds to putative proteins of 239-311 amino acid residues, respectively. All nine sequences, reconstructed from phylogenetic data showed sequences similar to the gamma-gliadins registered at GenBank. All known gamma-gliadin sequences fall into four major groups in the tree constructed by the neighbour-joining method. Primer. pairs were designed to discriminate between these four gamma-gliadin groups during grain development in bread wheat using quantitative real time polymerase chain reaction (QRT-PCR). Expression profiles of two lines of bread wheat showed higher levels of gene transcription for gamma-gliadins from group II, whereas gamma-gliadins from group I were expressed at the lowest level. Thus, the gamma-gliadins from group II are expressed four- and 10-fold higher in 'Perico' and 'Anza', respectively, than the other gamma-gliadins. The results reported may help to understand the order of synthesis and deposition of y-gliadins in the grain of developing bread wheat. In addition, the differences found in the expression levels among the four groups could be used to select genotypes with specific profiles of y-gliadins. These findings will also facilitate the up- and down-regulation of gamma-gliadin gene expression in transgenic wheat. (c) 2005 Elsevier Ltd. All rights reserved.