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分析丹参转录组数据库(SRX021907),得到一条R2R3-MYB基因,blast比对发现该基因为丹参Sm MYB7(KF059361.1)。分别从g DNA和c DNA水平克隆该基因全长,测序结果表明该基因与公布序列一致,分析发现该基因无内含子序列,包含一个长为954 bp的开放读码框(ORF),编码317个氨基酸残基。多重序列比对和系统进化树分析显示Sm MYB7蛋白与拟南芥At MYB73同属R2R3-MYB第22个亚家族。已有丹参基因信息结合BD walking获得1 974 bp的启动子序列,分析结果表明多种顺式作用元件存在于该基因的启动子区。实时荧光定量PCR分析显示,该基因的表达为组成型,在丹参根、茎、叶、花中都表达;随着花的发育,该基因的表达量逐渐增加;此外,Sm MYB7在盐胁迫、水杨酸、茉莉酸甲酯和脱落酸处理下表达均上调,推测该基因参与调控植物防御和花的发育。
Analysis of Danshen transcriptome database (SRX021907), to obtain a R2R3-MYB gene, blast comparison found that the gene is Salvia miltiorrhiza MYB7 (KF059361.1). The full length of this gene was cloned from g DNA and c DNA, respectively. The sequencing results showed that this gene was consistent with the published sequence. The gene was found to contain no intron sequence and contained a 954 bp open reading frame (ORF) 317 amino acid residues. Multiple sequence alignment and phylogenetic tree analysis showed that Sm MYB7 protein belongs to the 22nd subfamily of R2R3-MYB with Arabidopsis At MYB73. Danshen gene information has been combined with BD walking 1 974 bp promoter sequence analysis showed that a variety of cis-acting elements exist in the promoter region of the gene. Real-time PCR showed that the expression of Sm MYB7 was constitutively expressed in root, stem, leaf and flower of Salvia miltiorrhiza. The expression of Sm MYB7 increased with the development of flower. In addition, Salicylic acid, methyl jasmonate and abscisic acid were up-regulated, suggesting that the gene involved in the regulation of plant defense and flower development.