关键词: 多年生簇毛麦, a-醇溶蛋白基因, 基因克隆, 特异PCR

Abstract: PCR was carried out on the genomic DNA from Dasypyrum breviaristatum using the conserved primers specific forα-gliadin genes. The PCR products were cloned and sequenced. Total 7 sequences were obtained with length of 999 to 1 018 bp containing coding regions from 283 to 290 amino acids and their GenBank accession number are EU186102 to EU186108 respectively. BLAST search showed that these sequences belong to a-gliadin super-family. Searching for 4 different T cell stimulatory epitopes for celiac disease patient, it is found that 5 of 7 sequences contained the Glia-a-2 epitopes at the C terminal region. The phylogenetic trees indicated that all the sequences could not be clustered into the group of a-gliadin genes from the A, B, and D genomes of wheat (Triticum aestivum), but presented in a new group. On the basis of the a-gliadin gene sequences, a pair of AS-PCR primer was designed. PCR results indicated that the AS-PCR primers allow to specifically amplify the D. breviaristatum and wheat-D. breviaristatum partial amphiploid, which thus can be used as a marker to trace the D. breviaristatum derived a-gliadin genes in wheat-Dasypyrum introgression lines.

Key words: Dasypyrum breviaristatum, α-gliadin gene, Gene cloning, Specific PCR