小麦品系2004s-47的1Dx基因的克隆与序列分析

Molecular Cloning and Sequence Analysis of the 1Dx Gene in Wheat Line 2004s-47

明确2004s-47品系1Dx基因的序列,为寻找新的优质HMW-GS类型、实现小麦品质的改良及育种提供理论依据。应用SDS-PAGE对2004s-47品系中HMW-GS的亚基组成进行分析,用PCR技术克隆1Dx亚基基因并用DNAMAN软件进行序列分析。结果从2004s-47品系中扩增出了大小为2514 bp的基因。该基因具有典型的小麦HMW-GS x-型基因序列结构特征。又因为1D比1A和1B基因组中的x-型和y-型基因易表达,结合SDS-PAGE的结果,所以2004s-47品系中HMW-GS的亚基组成为(Null,7+8,?+10)。序列比对表明,2004s-47品系的1Dx?基因与节节麦(Aegilops tauschii)1Dx2.1t(AF480486)同源性最高。将该序列提交到NCBI,获得序列登陆号为:KP702118。2004s-47品系中1Dx?序列的确定,为原核表达确定1Dx?是否为新的基因提供了基础,也为品质改良及育种提供了依据。

Clarifying the sequence of 1Dx gene in 2004s-47 line would provide a theoretical basis for findingnew types of high-quality HMW-GS and achieve wheat quality improvement and breeding. The high molecularweight glutenin subunits（HMW-GS） composition of 2004s-47 line was analyzed by SDS-PAGE. 1Dx subunitgene was cloned by PCR and sequence analysis was performed by DNAMAN. The results revealed that thegene sequence that cloned from 2004s-47 was 2514 bp and had typical x-type structure of wheat HMW-GS.And because the expressions of x- and y-type genes in the D genome of bread wheat were easier than that inthe A and B genomes, together with the results of SDS-PAGE, the composition of 2004s-47 line was（Null,7＋8,？＋10）. Sequence alignments showed that the DNA sequence of the 1Dx？ gene from 2004s-47 had thehighest similarity to that of previously published 1Dx2.1t（AF480486） in Aegilops tauschii. The sequence wassubmitted to NCBI and sequence accession number： KP702118 was obtained. The identification of 1Dx genesequence in 2004s-47 line provided a theoretical basis for the determination of whether 1Dx？ was a new typeof gene by prokaryotic expression, and for wheat quality improvement and breeding.