关键词: 阿拉拉特小麦, 1Gy, 分子克隆, 原核表达, 序列分析

Abstract: In this study, genomic PCR reaction of Triticum araraticum(PI427305) was carried out by using degenerate primers specific for y-type HMW-GS.A 2.2 kb DNA fragment was amplified and cloned into the pMD18-T vector.DNA sequencing was carried out by using overlapping sub-clones.The complete coding region of the gene was 2 202 bp(GenBank accession number was HM131806), and 732 amino acid residues were encoded.It was found that the DNA sequence of this gene showed the highest identity with that of 1Gy7^* of Triticum timopheevii Zhuk, and the corresponding amino acid sequence had the typical structure of y-type HMW-GS.So it was concluded that this gene was 1Gy.The molecular weight of 1Gy was a little smaller than that of 1Gy7^*, and the mobility of 1Gy was slower than that of 1Gy7^*.Therefore, 1Gy was a novel y-type HMW-GS gene.Compared with other y-type subunits encoded by A, B and D genomes of common wheat, an extra cysteine existed in the repetitive domain near C-terminal domain of 1Gy.The repetitive domain was mainly proved to be coil by using PSIPRED.Both of them were beneficial to bread improvement.

Key words: Triticum araraticum, 1Gy, molecular cloning, prokaryotic expression, sequence analysis