摘要：

为明确光信号转导核心bZIP转录因子StHY5基因的基础特征与组织表达模式,以马铃薯栽培种底希瑞为试验材料,克隆了StHY5基因的全长CDS序列,通过生物信息学分析明确序列特征与进化关系,采用实时荧光定量PCR(qRT-PCR)技术分析在马铃薯不同组织中的表达模式。结果表明,StHY5蛋白由158个氨基酸组成,分子质量为17.32 ku,理论等电点为9.78,StHY5蛋白包含1个基础亮氨酸拉链(bZIP)结构域。StHY5蛋白的二级结构由α-螺旋(占比44.3%)和无规则卷曲(占比55.7%)构成,StHY5蛋白的三级结构以α-螺旋为主,参考模板为鸡CREB3 bZIP蛋白。系统进化树分析显示,StHY5蛋白与茄科植物的HY5蛋白聚为一支,且与辣椒CaHY5蛋白亲缘关系最近。qRT-PCR定量结果表明,StHY5基因在所有组织中均有表达,且表达量差异显著,其中,匍匐茎表达量最高、块茎表达量最低。明确了马铃薯StHY5基因的序列特征与组织表达模式,为后续开展该基因在光信号调控中的功能验证及作用路径分析提供了试验基础。

关键词: 马铃薯, StHY5, 基因克隆, 表达模式分析, 生物信息学分析, 光信号通路

Abstract:

To clarify the basic characteristics and tissue expression pattern of the light signal transduction core bZIP transcription factor StHY5 gene,using potato cultivar Désirée as the experimental material,the full-length CDS sequence of the StHY5 gene was cloned.The sequence characteristics and evolutionary relationship were clarified by bioinformatics analysis.The expression patterns in different potato tissues were analyzed by Quantitative Real-time PCR(qRT-PCR).The results showed that the StHY5 protein consisted of 158 amino acids,with a molecular weight of 17.32 ku and a theoretical isoelectric point of 9.78.It contained one basic leucine zipper(bZIP)domain.The secondary structure of StHY5 was composed of 44.3% α-helices and 55.7% random coils,while the tertiary structure was dominated by α-helices,using the chicken CREB3 bZIP protein as the template.Phylogenetic tree analysis revealed that StHY5 clusters with HY5 proteins from Solanaceae species,showing the closest genetic relationship to CaHY5 in capsicum.qRT-PCR quantification demonstrated that StHY5 was expressed in all tested tissues with significant expression differences,among them the highest expression in stolons,and the lowest expression in tubers.This study clarifies the sequence characteristics and tissue expression pattern of the potato StHY5 gene,providing an experimental foundation for subsequent functional verification and pathway analysis in light signal regulation.

Key words: Potato, StHY5, Gene cloning, Expression pattern analysis, Bioinformatics analysis, Light signaling pathway

中图分类号: 

• Q785转化及克隆