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本研究以三色堇(Viola×wittrockiana Gam)为研究材料,采用RT-PCR和RACE等方法克隆可能与其花色形成相关的酪氨酸脱羧酶基因(Tyrosine decarboxylase),并对基因全长进行氨基酸序列的生物信息学分析。结果从三色堇中分离克隆出c DNA全长为1 634 bp的TYDC1和1 575 bp的TYDC2两个同源基因,包含相同的且长度为1 485 bp,编码494个氨基酸的开放阅读框;氨基酸多重序列比对和聚类分析结果表明:三色堇中的TYDC基因编码氨基酸和其他植物相关基因的一致性可达到82.43%,且与麻疯树的亲缘关系较近,达到73%;对TYDC蛋白的结构进行预测发现,TYDC蛋白是一种亲水性的不稳定蛋白,也是一种存在跨膜区域且不存在信号肽的非分泌蛋白;通过亚细胞定位预测分析表明该蛋白主要在细胞核和细胞质中表达。本研究为揭示三色堇TYDC蛋白的结构功能务实了基础,为进一步研究和验证该基因在三色堇中的表达提供理论依据。
In this study, Viola × wittrockiana Gam was used as a material to clone the tyrosine decarboxylase gene possibly related to its flower color formation by RT-PCR and RACE. The amino acid sequence Bioinformatics analysis. Results Two orthologs of TYDC1 with a total length of 1 634 bp and TYDC2 with a length of 1 575 bp were isolated from Pansy and contained the same open reading frame with a length of 1 485 bp and encoding 494 amino acids. The results of amino acid sequence alignment and cluster analysis showed that the conformation of the TYDC gene in Pansy was 82.43%, which was close to that of Jatropha, reaching 73% TYDC protein structure prediction found that TYDC protein is a hydrophilic unstable protein, is also a transmembrane region and the absence of signal peptide non-secreted protein; by subcellular localization prediction analysis showed that the protein mainly in the nucleus And cytoplasmic expression. This study is to provide a solid foundation for revealing the structural function of TYDC and to provide a theoretical basis for further study and verification of its expression in Pansy.