马铃薯花青素转录激活基因stwd40的克隆与表达分析

Cloning and Expression Analysis of the Anthocyanin Transcriptional Activator Gene stwd40 of Solanum tuberosum

根据GenBank中报道的茄属植物矮牵牛花青素wd40类转录激活基因an11及番茄wd40基因113964R的mRNA序列的保守区结构,设计简并引物,从紫色马铃薯皮中克隆了马铃薯wd40类转录激活基因的保守片段,再利用RACE技术分别获得了该基因的3′端和5′端。序列分析表明,该基因核苷酸序列为1292bp,具有完整的编码框,推导其编码362个氨基酸,命名为stwd40。推测的氨基酸序列与GenBank数据库中大量物种的花青素转录激活蛋白wd40有较高的同源性,其中与矮牵牛花青素转录调控基因an11的相似性达86%。RT-PCR表达分析显示stwd40在紫色马铃薯叶、茎、皮、肉及根中都有表达,其中在茎中的表达量最高,肉中表达量最低;其在白色马铃薯的叶、皮和肉中也有表达,推测该基因为组成型表达。

Based on the conserved sequence of Petunia hybrida anthoeyain transcriptional activator gene wd40 （anl 1 ） and tomato wd40 mRNA （113964R） from GenBank, the conserved fragment of stwd40 transcriptional activator gene was cloned from the purple potato coat. The 3＇- end and 5＇- end of this gene were amplified by using RACE technique separately. Sequence analysis showed that the nucleotide sequence of this gene is 1 292 bp, containing a complete open reading frame and encoding 326 amino acids. The stwd40 amino acid sequence is similar to anthocyanin transcriptional activator protein wd40 from large number of species, and the homology is 86% with the Petunia hybrida anthocyain transcriptional regulatory gene anll. The results of RT-PCR expression analysis showed that stwd40 expressed in leaves, stems, tuber skins, tuber fleshes and roots of purple potato and the leaves, tuber skins, tuber fleshes of white potato with different expression levels. It is suggested that the stwd40 is constitutive expressed in potato.