甘菊内参基因ClTUA的克隆与表达分析  

Isolation and expression analysis of a reference gene:ClTUA of Chrysanthemum lavandulifolium.

作　　者：黄河[1] 牛雅静[1] 杨可[1] 戴思兰[1] 

机构地区：[1]北京林业大学园林学院

出　　处：《北京林业大学学报》2012年第2期112-117,共6页Journal of Beijing Forestry University

基　　金："863"国家高技术研究发展计划项目(2007AA021403);林业公益性行业科研专项(200904050)

摘　　要：根据前期工作已获得的甘菊α-tubulin基因EST序列,使用RACE技术获得了该基因的全长序列,命名为ClTUA。生物信息学分析表明,ClTUA基因全长cDNA序列为1580bp,编码432个氨基酸残基。利用MEGA4.0软件对推测的氨基酸序列进行系统关系分析发现,其为植物中Ⅰ类α-tubulin基因。RT-PCR分析发现:ClTUA基因不仅在盐、干旱、冷、热、脱落酸和水杨酸各种非生物胁迫下稳定表达,而且在不同生长发育阶段的样本中稳定表达;而作为对照的两个内参基因,ClActin和26SrRNA基因在不同非生物胁迫下表达稳定,但在不同生长发育阶段的样本中表达强度不同。这一结果表明,ClTUA基因的表达稳定性优于ClActin和甘菊26SrRNA基因,可以作为内参基因用于甘菊抗逆性和生长发育过程中基因表达规律研究。Selection and validation of reference genes constitute a key point for gene expression analysis of Chrysanthemum lavandulifolium. An EST encoding a-tubulin from C. lavandulifolium was identified in our previous study. By using RACE approach, we isolated the full length sequence of this gene and designated it as ClTUA. ClTUA contained an open reading frame of 1 580 bp that encoded a protein of 432 amino acids. Phylogenetic analysis using MEGA4.0 software indicated that ClTUA belonged to class I of a-tubulin genes. To assess the usability of ClTUA as a reference gene, we compared the expression patterns of CITUA with two other housekeeping genes, ClActin and 26S rRNA under salt, drought, cold, heat, ABA, and SA treatments as well as in different developmental stages. Our results showed that the transcript levels of ClActin and 26S rRNA were stable under abiotic treatments but varied in different developmental stages, while that of ClTUA was expressed consistently in all samples. The data suggest that ClTUA is more suitable than ClActin and 26S rRNA as a reference gene for the analysis of gene expression patterns under abiotic stress and in different growth stages in C. lavandulifolium.

关 键 词：甘菊 内参基因 C1TUA 基因表达分析 

分 类 号：S852.659.6[农业科学—基础兽医学]