小麦HMW-G12亚基基因启动子克隆及序列分析

Cloning and Sequencing Analysis of the Promoter for a High Molecular Weight Glutenin 12 Subunit gene from Wheat

为了研究高分子量谷蛋白基因启动子在种子中的特异性表达 ,以小麦品种“东农 774 2”的基因组DNA为模板 ,根据已发表序列设计并合成引物 ,用PCR的方法克隆了小麦贮藏蛋白中高分子量谷蛋白 12亚基基因的上游调控序列。序列测定结果表明 :所克隆的启动子片段大小为 4 2 4bp与Thomspon报道的序列比较 ,同源性为 97.9% ,有 9个核苷酸发生了改变。推测的TATAbox位于 - 2 7— - 30bp ,Prolamin -box位于 - 175— - 181bp ,认为该元件可能与转录速率的调控有关。

Glutenin is the most important component in wheat grain storage protein.And the content of glutenin has closed relation to wheat quality; especially the high molecular weight glutenin subunit (HMW-GS) plays an important role in bread-making quality.For the purpose of studying the seed-specific expression of wheat grain storage protein gene, The upstream regulatory region of a high molecular weight glutenin 12 subunit gene was amplified from Cultivar NE7742genomic DNA by polymerase chain reaction.Sequencing analysis showed the cloned fragment contained 424 nucleotides and shared a homology of 97.9% with the sequence published on genebank.There are 9 bp differences when compared with the reported sequence.The putative TATA box was present at position -27 to -30bp.A putative Prolamin-box and two Prolamin-like-boxes were found,and it might be important for the quantitative regulation of HMW glutenin 12 subunit gene expression.