Cloning and Sequence Analysis of α-gliadin Gene from Aegilops tauschii×Secale cereale Amphiploids

Fifteen novel α-gliadin genes from Aegilops tauschii×Secale cereale amphiploids were amplified and cloned using conserved primers specific to wheat α-gliadin genes.A total of 15 sequences were obtained with lengths ranging from 843 to 897 bp,containing ORFs encoding peptides with 280 to 298 amino acids and GenBank accession numbers JQ029719,JQ046392 to JQ046405,respectively.A BLAST search showed that these sequences belonged to typical structural characters of α-gliadin genes reported previously.Comparative analysis of homologous proteins showed that they were members of the α-gliadin super-family,of which two sequences were synonymous mutations.Searching for four different T cell stimulatory epitopes for celiac disease patients,it was found that eight of fifteen sequences did not contain the Glia-α-2 and Glia-α-9 epitope typical feature.The phylogenetic trees based on the amino acids sequence of Secale and Aegilops indicated that the cloned gliadin genes clustered in the clade of Aegilops α-gliadin gene.