实时荧光定量PCR构建奶牛生殖系统PrP基因标准品质粒和标准曲线  

Construction of cow reproductive systemPrPgene standard plasmid and curve using real-time RT-PCR

作　　者：张太翔[1] 宁章勇[2] 赵德明[1] 周向梅[1] 杨建民[1] 尹晓敏[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 [2]华南农业大学动物医学院,广州510642

出　　处：《中国农业大学学报》2007年第2期1-4,共4页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目(3057139930500371);教育部博士点基金资助项目(20050019031)

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。The purpose of this study was to elucidate the normal PrP gene expression and its normal physiological functions, 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 system PrP gene standard plasmid and curve using real-time RT-PCR. The result of the EcoR I to the plasmid including the target gene showed the product was 302 bp, which was according to the expected result. The constructed standard curve had a relative coefficient r^2 = 0.999 and the regressing equation= y = - 0.29x ＋ 9.48 suggested it was successful. The constructed calibration curve was used to quantify the unknown samples of cow.

关 键 词：PRP RT—PCR 实时荧光定量PCR 标准曲线 奶牛生殖系统 BSE 

分 类 号：S855.99[农业科学—临床兽医学]