葡萄风信子MaANS基因及启动子的克隆与分析

Isolation and Analysis of Muscari armeniacum MaANS Gene and Its Promoter

该研究根据转录组测序结果,在葡萄风信子（Muscari armeniacum）‘亚美尼亚’中克隆到花青素合酶（ANS）基因的cDNA与DNA序列,该基因命名为MaANS。采用荧光实时定量分析MaANS时空表达模型,同时利用染色体步移法克隆到MaANS上游1 044bp的一段序列。信息学分析表明：MaANS开放阅读框为1 065bp,编码355个氨基酸;DNA与cDNA的一致性为89.62%,DNA序列在ATG下游515~594bp之间插入1个79bp内含子;启动子序列在-70bp位置有1个TATA-box,有多个光响应元件及MYB结合位点等。荧光实时定量分析表明,MaANS基因在花中优势表达,并且在完全着色期表达量最高。该研究结果为深入研究MaANS基因功能、分析葡萄风信子着色机理奠定了基础。

In this study,we cloned cDNA and DNA sequences of anthocyanin synthase（ANS）gene,named MaANS,based on transcriptome sequencing in Muscari armeniacum.We did real-time fluorescence quantitative analysis about MaANS.While taking advantage of chromosome walking.We cloned a 1 044 bp upstream sequence of MaANS.Bioinformatic analysis showed that MaANScontains a 1 065 bp open reading frame,encoding 355 amino acids.The consistency of cloned DNA and cDNA is 89.62%,DNA sequence inserts a 79 bp intron from 515 bp to 594 bp on downstream of ATG.TATA-box is at-70 bp of the promoter which has a plurality of light-responsive elements and MYB binding sites.Real-time fluorescence quantitative analysis showed MaANSwas predominant expression in flower and the expression level at fully colored stage was the highest.In this study,gene cloning and expression analysis on MaANSare important to in-depth study the function of MaANSgene and analyze the colored mechanism of Muscari armeniacum.