小麦‘Fielder’ ω-醇溶蛋白基因及启动子的克隆与序列分析

Cloning and sequence analysis of the coding region and promoter of ω-gliadin genes from wheat ‘Fielder'(Triticum aestivum L.)

关键词:  小麦  A-PAGE  醇溶蛋白  启动子  endosperm box

Key words:  wheat  A-PAGE  gliadin  promoter  endosperm box

摘要:

为研究六倍体小麦中ω-醇溶蛋白基因簇的启动子差异，以普通小麦品种‘Fielder’为试验材料，利用同源克隆的方法，克隆ω-醇溶蛋白基因的编码区和启动子序列，并进行生物信息学相关分析。结果表明:1)共克隆得到11条不同的基因序列(MN441496~MN441506)，其中6条(MN441496~MN441497和MN441503~MN441506)编码ARE/Q型ω-醇溶蛋白，5条(MN441498~MN441502)编码SRL型ω-醇溶蛋白。2)ARE/Q型ω-醇溶蛋白基因编码区长度范围在972~1 158 bp，SRL型ω-醇溶蛋白基因编码区长度范围在1 303 ~1 419 bp，这种差异主要由中间重复区的Indel类型和数量不同造成。3)ARE/Q型ω-醇溶蛋白基因启动子在-300 bp含有1个典型的endosperm box，而SRL型ω-醇溶蛋白基因启动子在-300和-600 bp处各含有1个endosperm box。这些ω-醇溶蛋白基因启动子上的差异，可能引起表达水平的差异。

Abstract:

To understand the promoter differences of ω-gliadin gene cluster in hexaploid wheat, the coding region along with the promoter sequences of ω-gliadin in hexaploid common wheat cultivar ‘Fielder' were obtained by homologous cloning. Bioinformatics analysis was conducted. The results showed that: 1)A total of 11 different ω-gliadin gene sequences were acquired, and six of which belonged to the ARE/Q type genes, the rest five were SRL type genes. 2)The coding region lengths were varied from 972 to 1 158 bp among the six ARE/Q type genes and from 1 303 to 1 419 bp among the five SRL type genes, which were the results of differences in types and quantities of Indels in the center repeat region. 3)There located a conserved typical endosperm box at -300 bp upstream of ARE/Q type ω-gliadin genes. Nevertheless, the endosperm box appeared at -300 and -600 bp upstream of SRL type ω-gliadin genes, respectively. The differences in the promoter of ω-gliadin gene cluster might influence the expression levels of different ω-gliadin genes.