橡胶草TkBZR家族基因的鉴定与原核表达分析

Genome-wide identification and prokaryotic expression analysis of TkBZR family genes in rubber dandelion (Taraxacum kok-saghyz)

摘要

为了进一步了解TkBZRs基因在橡胶草（Taraxacum kok-saghyz）生长发育和抵抗逆境中的作用，本研究通过分析橡胶草的全基因组测序数据，对TkBZR/BES家族基因进行鉴定分析，得到了6个TkBZR/BES家族成员，分别命名为TkBZR1至TkBZR6，并探讨其在染色体上的分布、结构域特征、组织特异性和时空表达特征。结果显示，6个TkBZR/BES家族成员分布在6条独立的骨架上。系统发育树显示，TkBZR/BES基因家族成员在进化上保守，橡胶草与莴苣的基因聚类在同一亚组，表明两者之间可能存在较近的亲缘关系。基因结构和保守结构域分析结果显示，除了TkBZR5外，其余TkBZR基因都包含2个外显子和1个内含子，所有成员都展示了高度保守的BES1_N结构域。表达模式分析发现，有5个成员在所有5个组织中表达，1个成员几乎不表达。此外，成功克隆获得表达丰度最高的TkBZR2基因，该基因编码307个氨基酸。将TkBZR2同源重组至原核表达载体上，重组成功后转入大肠杆菌（Escherichia coli）BL21(DE3)中并表达TkBZR2重组蛋白。

Abstract

To further understand the role of the TkBZR family genes in the growth, development, and stress resistance of rubber dandelion (Taraxacum kok-saghyz), the TkBZR/BES family genes were identified and analyzed by examining the whole genome sequencing data of T. kok-saghyz. A total of 6 members of the TkBZR family were identified, named from TkBZR1 to TkBZR6, and their distribution on chromosomes, domain characteristics, tissue expression profiles and the spatial-temporal gene expression were analyzed. The results revealed that TkBZR/BES gene family members were distributed on 6 independent scaffolds. Phylogenetic trees indicate thatTkBZR/BES gene family members are evolutionarily conserved, with rubber dandelion and lettuce genes clustering in the same subgroup, suggesting a possible close evolutionary relationship between the two. Gene structure and conserved domain analysis show that except for TkBZR5, all the other TkBZR genes contain two exons and one intron, and that all the family members exhibit a highly conserved BES1_N domain. Expression pattern analysis reveals that 5 members are expressed in all five tissues, while one member is almost unexpressed. Additionally, the TkBZR2 gene with the highest expression abundance was successfully cloned, encoding 307 amino acids. Homologous recombination of TkBZR2 was made into prokaryotic expression vector, and the recombinant plasmid was transferred into E. coli BL21 (DE3). The recombinant protein TkBZR2 was expressed in E. coli BL21 (DE3).

关键词

橡胶草 / BZR/BES / 基因家族 / 基因结构分析 / 原核表达

Key words

Taraxacum kok-saghyz / BZR/BES / gene family / gene structure analysis / prokaryotic expression

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基金资助

“崖州湾”菁英人才科技专项项目(SCKJ-JYRC-2023-19)

海南省重点研发项目(ZDYF2022XDNY251)

国家自然科学基金项目(32260400)