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近年来实时荧光定量PCR已经成为研究基因表达的标准方法,稳定的内参基因可使反应标准化进行,并提高该方法的敏感度和重复性。许多研究表明不同组织、细胞和不同条件下内参基因的表达存在很大差异,因此在研究基因表达分析时,需要以内参基因对目标基因的表达量进行校准,由此来获得更为准确的结果。本研究以番茄经过果糖、蔗糖、葡萄糖、高温和低温处理的材料为研究对象,利用实时荧光定量PCR方法,对Actin、CAC、TIP41、Expressed、SAND 5个内参基因m RNA水平的表达量进行分析。经ge Norm和Norm Finder软件分析5个内参基因的表达稳定性,以期为番茄果实基因表达调控相关的研究提供内参基因。研究结果为番茄植株在非生物胁迫下的实时定量RT-PCR分析中内参基因的选择提供了理论与实验依据。
In recent years, real-time fluorescence quantitative PCR has become the standard method for studying gene expression. The stable internal control gene can standardize the reaction and improve the sensitivity and repeatability of the method. Many studies show that there are great differences in the expression of internal reference genes in different tissues, cells and under different conditions. Therefore, when analyzing gene expression analysis, it is necessary to calibrate the expression level of the target gene with the internal reference gene to obtain more accurate results . In this study, we used tomato, fructose, sucrose, glucose, high temperature and low temperature materials to study the expression of Actin, CAC, TIP41, Expressed, SAND mRNA expression levels of five reference genes using real-time fluorescence quantitative PCR . The gene expression stability of five internal control genes was analyzed by ge Norm and Norm Finder software to provide reference genes for the research related to the regulation of gene expression in tomato fruit. The results provided theoretical and experimental basis for the selection of reference genes in real-time quantitative RT-PCR analysis of tomato plants under abiotic stress.