作 者：
韩彩霞;赵德明;吴长德;宁章勇;杨建民;刘美丽;马李颖
单 位：
中国农业大学动物医学院国家动物海绵状脑病实验室
关键词：
PrP;RT-PCR;实时荧光定量PCR;标准曲线;绵羊
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.
译 名：
Construction of Sheep PrP Gene Standard Plasmid DNA and Curve Using Real-time RT-PCR
作 者：
HAN Cai-xia,ZHAO De-ming~*,WU Chang-de,NING Zhang-yong,YANG Jian-min,LIU Mei-li,MA Li-ying(National Laboratory for Transmissible Spongiform Encephalopathy,College of VeterinaryMedicine,China Agricultural University,Beijing 100094,China)
关键词：
PrP;RT-PCR;real-time quantitative PCR;standard curve;sheep
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.
