实时荧光定量PCR中内参基因的选择.pdfVIP

微生物学通报 DEC 20, 2010, 37(12): 1825−1829 Microbiology China © 2010 by Institute of Microbiology, CAS tongbao@ 生物实验室 实时荧光定量 PCR 中内参基因的选择 赵文静 徐洁 包秋华 陈永福 张和平* ( 内蒙古农业大学 乳品生物技术与工程教育部重点实验室 内蒙古 呼和浩特 010018) 摘 要: 实时荧光定量PCR 技术是分析基因表达谱的一种常用方法, 在分析中选择合适的内参基 因对数据进行校正是得到可信数据的关键。以Lactobacillus helveticus H9 为研究对象, 应用实时 荧光定量PCR 技术, 评价了5 种常用内参基因ldh 、recA 、rpoB 、gapdh 和 16S rRNA 的表达稳定 性, 通过 geNorm 和NormFinder 程序进行数据分析, 结果表明5 个候选内参基因在菌株不同的发 酵时间点表达相对都较为稳定, 结合两种分析得到其中最为稳定的基因是ldh, 适合于用作后续实 时荧光定量PCR 试验中的内参基因。 关键词: 实时荧光定量PCR, 内参基因, GeNorm, NormFinder Selection of Reference Genes for Real-time Quantitative PCR ZHAO Wen-Jing XU Jie BAO Qiu-Hua CHEN Yong-Fu ZHANG He-Ping* (Key Laboratory of Dairy Biotechnology and Engineering , Ministry of Education , Inner Mongolia Agricultural University , Huhhot , Inner Mongolia 010018, China) Abstract: Real-time quantitative PCR is a commonly used method to analyse the gene expression pro- file, it is important to select an appropriate reference gene for normalization of experimental data when using this method. In our study, we used two statistical methods to evaluate the gene expression stabili- ties of five reference genes (ldh, recA, rpoB, gapdh and 16S rRNA) under the different growth phases of Lactobacillus helveticus H9. The results showed that the best reference gene was ldh which was the most stable gene would be used for normalization of real-time quantitative PCR experiments data. Keywords: Real-time quantitative PCR, R