根据小麦低分子量谷蛋白基因保守区序列设计引物P1/P2, 采用PCR法对四川小麦地方品种AS1643的基因组DNA进行扩增, 获得1条约900 bp的片段, 分离、纯化后连接到载体pMD18-T上, 对筛选阳性克隆测序, 获得1个低分子量谷蛋白基因LMW-AS1643(GenBank登录号: EF190322), 其编码区长度为909 bp, 可编码302个氨基酸残基组成的成熟蛋白。序列分析结果表明, LMW-AS1643具有典型的低分子量谷蛋白基因的基本结构, 其推导氨基酸序列与其它已知的LMW-GS相比, 最高相似性为93.40%。生物信息学分析表明, 在LMW-AS1643低分子量谷蛋白中, 无规则卷曲含量最高, 为67.90 %, 其次是a-螺旋, 占30.46 %, b-折叠含量最少, 为1.64 %。

The full-length coding region (open reading frame, ORF) of low-molecular-weight glutenin subunit (LMW-GS ) gene was amplified from Sichuan wheat landrace accession AS1643 by using the primer pairs P1/P2, which were designed according to wheat LMW-GS conservative domains. The amplified DNA fragment was separated and recovered from agarose gel, subsequently cloned into pMD18-T vector, and then transformed into E. coli strain DH5a. One positive clone LMW-AS1643 (GenBank accession No. EF 190322) was selected and sequenced. It had a coding region of 909 bp, and encoded a mature protein of 302 amino acid residues. Sequence analysis suggested that LMW-AS1643 was a typical LMW-GS gene. The deduced amino acid sequence comparison suggested that LMW-AS1643 had a high similarity with other known LMW-GS genes, with the highest similarity of 93.40 %Bioinformatics analysis indicated that among the three types of secondary structures, content of coil was the highest with 67.90 %, a-helix was the second with 30.46%, and b-sheet was the lowest with 1.64%.