Molecular characterization of α-gliadin genes from common wheat cultivar Zhengmai 004 and their role in quality and celiac disease

作者：Yuge Li, Ranran Xin, Dale Zhang, Suoping Li · 发表于：The Crop Journal · 年份：2013 · DOI：10.1016/j.cj.2013.11.003 · 被引用次数：22 · 研究领域：Celiac Disease Research and Management、Wheat and Barley Genetics and Pathology、Galectins and Cancer Biology

A total of 43 unique clones (Z4A-1 to Z4A-43 with GenBank accession numbers of HM120221, HM120222, JX828270, JN831402 to JN831406JN831402JN831403JN831404JN831405JN831406, and KC715889 to KC715923KC715889KC715890KC715891KC715892KC715893KC715894KC715895KC715896KC715897KC715898KC715899KC715900KC715901KC715902KC715903KC715904KC715905KC715906KC715907KC715908KC715909KC715910KC715911KC715912KC715913KC715914KC715915KC715916KC715917KC715918KC715919KC715920KC715921KC715922KC715923, respectively) were amplified and cloned from common wheat cultivar Zhengmai 004 using a PCR-based strategy. They included 22 full-ORF α-gliadin genes and 21 pseudogenes containing at least one in-frame stop codon. Comparative analysis of the deduced amino acid sequences showed that all the isolated genes displayed the typical structural features of α-gliadin genes and that the putative proteins of Z4A-7, Z4A-14, Z4A-17 and Z4A-20 had an extra cysteine residue in the unique domain II, while Z4A-15 lacked the second conserved cysteine residue in the unique domain I. The two fusion proteins of Z4A-15 and Z4A-20 were successfully detected by SDS-PAGE and Western blotting, although the protein level was relatively low. Based on the occurrence of the four major epitopes, as well as the lengths of the two glutamine repeats, 8, 6, and 8 genes were assigned to the Gli-2 loci on the respective chromosomes 6A, 6B, and 6D and a total of respectively 16, 0 and 23 immunogenic peptides were identified. In addition, 4 of th...