实时荧光定量PCR构建绵羊PrP基因标准品质粒和标准曲线  

Construction of Sheep PrP Gene Standard Plasmid DNA and Curve Using Real-time RT-PCR

作　　者：韩彩霞[1] 赵德明[1] 吴长德[1] 宁章勇[1] 杨建民[1] 刘美丽[1] 马李颖[1] 

机构地区：[1]中国农业大学动物医学院国家动物海绵状脑病实验室,北京100094

出　　处：《畜牧兽医学报》2005年第12期1363-1366,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(3037106230400325);教育部博士点基金(20020019006)

摘　　要：Here we report the construction of sheep PrP gene standard plasmid DNA and curve using real-time RT-PCR.Total RNA was extracted from each sample and the fragments of target gene were amplified by RT-PCR.The plasmid was constructed for calibrating unknown samples.In this study,the constructed plasmid containing only the target gene was used to construct a calibration curve.The absolute standard curve method was shown to be of high linearity,sensitivity and reproducibility.The purpose of this study is to investigate the quantification of PrP mRNA expression for knowing the scrapie pathogenesis and providing powerful tool for further studies on prion diseases pathogenesis.Here we report the construction of sheep PrP gene standard plasmid DNA and curve using realtime RT-PCR. Total RNA was extracted from each sample and the fragments of target gene were amplified by RT-PCR. The plasmid was constructed for calibrating unknown samples. In this study, the constructed plasmid containing only the target gene was used to construct a calibration curve. The absolute standard curve method was shown to be of high linearity, sensitivity and reproducibility. The purpose of this study is to investigate the quantification of PrP mRNA expression for knowing the scrapie pathogenesis and providing powerful tool for further studies.

关 键 词：PRP RT—PCR 实时荧光定量PCR 标准曲线 绵羊 

分 类 号：S852.659.7[农业科学—基础兽医学]