Cloning, expression, and evolutionary analysis of α-gliadin genes from Triticum and Aegilops genomes

J. Li, S. L. Wang, M. Cao, D. W. Lv, S. Subburaj, X. H. Li, F. J. Zeller, S. L.K. Hsam, Y. M. Yan

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Fifteen novel α-gliadin genes were cloned and sequenced from Triticum and related Aegilops genomes by allele-specific polymerase chain reaction (AS-PCR). Sequence comparison displayed high diversities in the α-gliadin gene family. Four toxic epitopes and glutamine residues in the two polyglutamine domains facilitated these α-gliadins to be assigned to specific chromosomes. Five representative α-gliadin genes were successfully expressed in Escherichia coli, and their amount reached a maximum after 4 h induced by isopropyl-β-D-thiogalactoside (IPTG), indicating a high level of expression under the control of T7 promoter. The transcriptional expression of α-gliadin genes during grain development detected by quantitative real-time polymerase chain reaction (qRT-PCR) showed a similar up-down regulation pattern in different genotypes. A neighbor-joining tree constructed with both full-open reading frame (ORF) α-gliadin genes and pseudogenes further revealed the origin and phylogenetic relationships among Triticum and related Aegilops genomes. The evolutionary analysis demonstrated that α-gliadin genes evolved mainly by synonymous substitutions under strong purifying selection during the evolutionary process.

Original languageEnglish (US)
Pages (from-to)157-167
Number of pages11
JournalJournal of Applied Genetics
Volume54
Issue number2
DOIs
StatePublished - May 2013
Externally publishedYes

Keywords

  • Heterologous expression
  • Phylogenetic evolution
  • Quantitative real-time PCR
  • Toxic epitopes
  • α-Gliadins

ASJC Scopus subject areas

  • Genetics

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