金钗石斛5个管家基因序列和表达分析——基因表达定量中内参基因的选择

Analysis of Sequences & Expression of Five House-keeping Genes in Dendrobium nobile——Selection of Reference Genes for Quantitative Property of Gene Expression

基因表达分析如定量PCR、RNA印记等需要应用内参基因对目标基因表达量进行校正,以期获得真实可靠的结果,管家基因是常用的内参基因。热带兰金钗石斛中管家基因的研究至今尚无报道。该文对金钗石斛腋芽cDNA文库中的5个管家基因的序列和基因表达进行了分析和检测。其中,4个基因编码泛素蛋白,1个编码a-微管蛋白,分别命名为DnUBQ1、DnUBQ2、DnUBQ3、DnUBQ4及DnTUA1.DnUBQ1基因的密码使用频率表现出与金钗石斛普遍密码子不同的偏好性。在低温春化前后和春化过程中,5个基因的表达均无明显改变,表现为组成型表达。因此,5个基因均可在研究金钗石斛春化相关基因的表达时作为参照基因使用。

House-keeping genes often serve as internal references in quantitation of gene expression level.To validate the reliability of five house-keeping genes in Dendrobium nobile,bioinformatics analysis on cDNA sequences and their expression under vernalization were conducted in this study.Among the five genes,BlastX searching indicated that 4 coded for homologous proteins of Ubiquitins while the left one encoded α-tubulin protein.Expression analysis showed no obvious changes occurred after and during the vernalization,suggesting that these genes would be reliable as the internal control in quantitation of genes expression in the process of vernalization.