植物实时荧光定量PCR内参基因的特点及选择

Characterization and Selection of Reference Genes for Real-time Quantitative RT-PCR of Plants

实时荧光定量PCR(qRT-PCR)具有灵敏度高、特异性强、重复的动态定量范围和高通量等优点,是进行植物基因表达和转录分析最常用的技术手段之一。选择合适的内参基因是正确运用实时荧光定量PCR分析目标基因表达变化的前提。近年来,大量研究表明,内参基因的选择应取决于研究者的实验条件;随着实验条件的变化,内参基因的选择也随之变化。因此,实时荧光定量PCR结果分析的准确性在很大程度上依赖于所选择的内参基因是否适合。该文从内参基因的选择、常用内参基因的特点、新内参基因的挖掘、应用内参基因组合的优点和内参基因的稳定性评价等几方面进行综述,以期为研究者在实验中选择合适的内参基因提供参考和理论依据。

Real-time quantitative RT-PCR （qRT-PCR） is one of the most common technologies used for gene expression and transcriptome analysis, with its high sensitivity, specificity, good reproducibility, wide dynamic quantification range and high-throughput capacity. Selecting the appropriate reference genes is the first step in analyzing the expression of genes of interest. Selecting appropriate reference genes depends on experimental conditions, and selection of reference genes changes after the experimental conditions. Therefore, the accuracy of results from qRT-PCR analysis largely de- pends on the reference genes used. In this paper, we give a comprehensive summary of reference genes for qRT-PCR, including their selection, characteristics of traditional reference genes, mining new reference genes, the advantage of combining different reference genes, and how to assess stable reference gene expression. The results provide a theo- retical foundation for selecting appropriate reference genes for qRT-PCR of plants.