作 者：
张太翔;宁章勇;赵德明;周向梅;杨建民;尹晓敏
单 位：
华南农业大学动物医学院;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;奶牛生殖系统;BSE
摘 要：
为阐释PrP基因在奶牛生殖系统的正常表达和正常的生理功能,以及为确定生殖系统在疯牛病垂直传播中的地位和其分子机理奠定基础,采用实时荧光定量RT-PCR的方法构建了奶牛生殖系统PrP基因的标准质粒和标准曲线。对含有目的基因质粒的EcoRⅠ酶切鉴定表明所插入的片段大小为302 bp,与预期的结果一致。所构建的标准曲线的相关系数r2=0.999,系统生成的回归方程为y=-0.29x+9.48,说明成功构建了奶牛生殖系统PrP基因的标准质粒和标准曲线,为研究PrP基因在奶牛其他组织的定量奠定基础。
译 名：
Construction of cow reproductive systemPrPgene standard plasmid and curve using real-time RT-PCR
作 者：
Zhang Taixiang1,Ning Zhangyong2,Zhao Deming1,Zhou Xiangmei1,Yang Jianmin1,Yi Xiaomin1(1.National Laboratory of Animal Transmissible Spongiform Encephalopathies,College of Veterinary Medicine,China Agricultural University,Beijing100094,China;2.College of Veterinary Medicine,South China Agricultural University,Guangzhou510642,China)
关键词：
PrP;RT-PCR;real-time quantitative PCR;standard curve;cow reproductive system;BSE
摘 要：
The purpose of this study was to elucidate the normalPrPgene expression and its normal physiological func-tions,confirm the role of the reproductive system in the maternity transmission route of the BSE and establish a good foundation for further molecular studies on BSE spread.Here we report the construction of cow reproductive systemPrP gene standard plasmid and curve using real-time RT-PCR.The result of theEcoRⅠto the plasmid including the target gene showed the product was302bp,which was according to the expected result.The constructed standard curve had a relative coefficientr2= 0.999and the regressing equation:y=-0.29x+ 9.48suggested it was successful.The constructed calibration curve was used to quantify the unknown samples of cow.
