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Molecular characterization of the celiac disease epitope domains in α-gliadin genes in Aegilops tauschii and hexaploid wheats (Triticum aestivum L.)

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Abstract

Nineteen novel full-ORF α-gliadin genes and 32 pseudogenes containing at least one stop codon were cloned and sequenced from three Aegilops tauschii accessions (T15, T43 and T26) and two bread wheat cultivars (Gaocheng 8901 and Zhongyou 9507). Analysis of three typical α-gliadin genes (Gli-At4, Gli-G1 and Gli-Z4) revealed some InDels and a considerable number of SNPs among them. Most of the pseudogenes were resulted from C to T change, leading to the generation of TAG or TAA in-frame stop codon. The putative proteins of both Gli-At3 and Gli-Z7 genes contained an extra cysteine residue in the unique domain II. Analysis of toxic epitodes among 19 deduced α-gliadins demonstrated that 14 of these contained 1–5 T cell stimulatory toxic epitopes while the other 5 did not contain any toxic epitopes. The glutamine residues in two specific ployglutamine domains ranged from 7 to 27, indicating a high variation in length. According to the numbers of 4 T cell stimulatory toxic epitopes and glutamine residues in the two ployglutamine domains among the 19 α-gliadin genes, 2 were assigned to chromosome 6A, 5 to chromosome 6B and 12 to chromosome 6D. These results were consistent with those from wheat cv. Chinese Spring nulli-tetrasomic and phylogenetic analysis. Secondary structure prediction showed that all α-gliadins had high content of β-strands and most of the α-helixes and β-strands were present in two unique domains. Phylogenetic analysis demonstrated that α-gliadin genes had a high homology with γ-gliadin, B-hordein, and LMW-GS genes and they diverged at approximate 39 MYA. Finally, the five α-gliadin genes were successfully expressed in E. coli, and their expression amount reached to the maximum after 4 h induced by IPTG, indicating that the α-gliadin genes can express in a high level under the control of T7 promoter.

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Acknowledgments

This research was financially supported by grants from the National Natural Science Foundation of China (30830072, 30771334), the Chinese National Basic Research Program (2009CB118303), National 863 project (2006AA10Z186), and Beijing Natural Scientific foundation and the Key Developmental Project of Science and Technology of Beijing Municipal Commission of Education (KZ200910028003).

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Authors and Affiliations

  1. Key Laboratory of Genetics and Biotechnology, College of Life Science, Capital Normal University, Beijing, 100048, China

    Zhenze Xie, Congyan Wang, Ke Wang, Shunli Wang, Xiaohui Li, Zhao Zhang & Yueming Yan

  2. Faculty of Life Science, Langfang Teachers College, Langfang, 065000, Hebei, China

    Congyan Wang

  3. Western Australian Department of Agriculture and Food, State Agriculture Biotechnology Centre, Murdoch University, Perth, WA, 6150, Australia

    Wujun Ma

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  1. Zhenze Xie
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  2. Congyan Wang
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  3. Ke Wang
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  4. Shunli Wang
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  5. Xiaohui Li
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  6. Zhao Zhang
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  7. Wujun Ma
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Correspondence to Wujun Ma or Yueming Yan.

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Communicated by B. Friebe.

Z. Xie, C. Wang, and K. Wang contributed equally to this work.

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Xie, Z., Wang, C., Wang, K. et al. Molecular characterization of the celiac disease epitope domains in α-gliadin genes in Aegilops tauschii and hexaploid wheats (Triticum aestivum L.). Theor Appl Genet 121, 1239–1251 (2010). https://doi.org/10.1007/s00122-010-1384-8

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