反转录实时定量PCR在植物基因表达上的进展_胡昊技术总结.pdfVIP

中国农学通报 2013,29(15): 127-134 Chinese Agricultural Science Bulletin 反转录实时定量PCR 在植物基因表达分析上的研究进展 胡 昊，李进进，王彩云 （华中农业大学园艺植物生物学教育部重点实验室，武汉430070 ） 摘 要：近年来，反转录实时定量PCR 作为一项新的核酸序列定量分析技术以其定量准确、高灵敏度和 高通量等特点，已被广泛的应用于分子诊断、病理分析以及食品安全检测等方面。然而其在植物基因表 达水平研究上的报道较为少见。在前人研究基础上，总结了反转录实时定量PCR 在植物基因综合表达 分析、基因家族表达分析、基因差异表达分析、转基因表达分析等方面的应用；分析了反转录、看家基因 选择等影响检测基因表达的因素；指出该技术在重复性、灵敏度、标准化上所存在的问题。随着分子生 物技术的发展，反转录实时定量PCR 将会在植物遗传育种、植物病理检疫以及分子进化等方面发挥更 为广泛的作用。 关键词：实时定量PCR ；植物基因表达；反转录；看家基因 中图分类号：Q789 文献标志码：A 论文编号：2012-2632 Research Advances of Real-time Quantitative Reverse Transcription PCR in Plant Gene Expression Hu Hao, Li Jinjin, Wang Caiyun (KeyLaboratoryofHorticulturalPlantBiology,MinistryofEducation,HuazhongAgricultural University, Wuhan 430070) Abstract: In recent years, the real-time transcription polymerase chain reaction (RT-PCR) has been widely used to molecular diagnostics, pathological analysis and food safety test as a new methodology of nucleic acid sequences quantitative analysis. Although the RT-PCR was a quantitative accurate, highly sensitive and high-throughput technology, its study on plant gene expression was relatively few. The article summarized the applications of RT-PCR in analysis of gene integrated expression, gene families expression, gene differential expression and transgenic expression. Based on case studies, the influencing factors of gene expression detection were analyzed by including reverse transcription and housekeeping gene selection. However, the technology still needed to be improved due to its reproducibility, sensitivity and normalization flaws. With the development of molecular biotechnology, the RT-PCR will play a more extensive role i