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[目的]克隆元宝枫转录因子MYB基因,为进一步研究元宝枫MYB基因功能和对其花青苷结构靶基因的调控研究奠定基础。[方法]以元宝枫‘鲁红1号’为材料,采用RT-PCR和RACE-PCR方法,克隆元宝枫‘鲁红1号’中MYB基因。[结果]测序结果显示该基因全长831 bp,编码276个氨基酸,该蛋白分子量为32.17 kDa,分子式为C_(1430)H_(14052)N_(2247)O_(406)S_(14),原子总数为4 510个,等电点为9.44,GenBank登录号为1825712,命名为AtrMYB。该蛋白具有R2R3MYB结构域,该蛋白偏疏水性,没有信号肽,具有核定位信号。氨基酸序列比对发现与其它物种的MYB有较高的同源性,进化树分析表明,AtrMYB与调控橙子花青苷合成的转录因子MYB亲缘关系最近,处在同一进化枝。[结论]从元宝枫‘鲁红1号’中成功获得了元宝枫转录因子MYB基因。
[Objective] The aim of this study was to clone the MYB gene of Acer truncatum and lay the foundation for the further study on MYB gene function and regulation of its anthocyanin structural target genes. [Method] With RT-PCR and RACE-PCR method, the MYB gene in Yuanhong Maple ’Luhong 1’ was cloned by RT-PCR and RT-PCR. [Result] The result of sequencing showed that the gene was 831 bp in length and encoded 276 amino acids. The molecular weight of this gene was 32.17 kDa and the molecular formula was C 1430 H 14041 N 2247 O 406 H 14. The total number of atoms 4 510, isoelectric point 9.44, GenBank accession number 1825712, named AtrMYB. This protein has the R2R3MYB domain, which is hydrophobic, has no signal peptide and has a nuclear localization signal. Amino acid sequence alignment showed high homology with MYB of other species. Phylogenetic tree analysis showed that AtrMYB had the closest relationship with MYB, a transcription factor regulating cyanidin synthesis, and was located in the same clade. [Conclusion] MYB gene of Acer truncatum was successfully obtained from ’Yuanhong 1’.