【摘要】 用SYBR GreenⅠ荧光实时定量PCR技术,以β-actin为参照基因,建立了猪肌肉中解偶联蛋白3(UCP3)基因mRNA丰度的定量分析方法。UCP3基因片段的PCR产物测序与GenBank中该基因的序列一致,定量PCR过程中UCP3的熔解曲线也表明该PCR过程为特异性扩增。猪肌肉样品UCP3 mRNA样品的批内和批间变异系数分别<2%和<11%,标准曲线R2=0.998,线性范围涵盖样品的Ct值。由此,实验建立的猪肌肉UCP3基因mRNA丰度测定方法是可靠的、准确的,可以用于猪肌肉中UCP3 mRNA丰度的测定。更多还原

【Abstract】 In this study,using SYBR Green Ⅰas a fluorescence dye,the real-time quantitative RT-PCR method was adapted to determine uncoupled protein-3(UCP3) mRNA abundance in the longissimus muscle of pigs with the reference gene of β-actin.Based on the specificity of RT-PCR of UCP3,linearity of standard cure,and intra and inter-assays CV being <2% and <11%,this established method is valid to assay the abundance of UCP3 mRNA in muscle of pigs.更多还原