鸡核受体辅激活蛋白1基因的荧光定量PCR检测

摘要

The expression of nuclear receptor co-activator 1 ( NC0A1) of chicken mRNA is key to development and maturity of gonads, and is related to reproduction performance. The aim of this study was to establish a fluorescent quantitative PCR (FQ-PCR) method for NC0A1 expression. The primer were designed and synthesized according to the NC0A1 sequence of chicken available in GenBank. The real-time RT-PCR assay was developed and assessed by reference gene of βaction. Results showed that the standard curve had good linear dependence, and the fluorescent quantitative RT-PCR assay was sensitive, specific and reliable. The results indicated that the relative expression of NCOA1 mRNA could be detected u-sing the real-time PCR with SYBR Green Ⅰ.%鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用，并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法，根据GenBank中NCOA1基因序列设计合成了引物，以β-action基因为内参基因，对荧光定量PCR方法进行了评估。结果表明：构建的标准曲线线性关系良好，建立的NCOA1基因荧光定量PCR检测方法灵敏度高、特异性强，准确可靠。建立的SYBR Green I实时荧光定量PCR方法可以测定鸡NCOA1 mRNA表达水平的相对含量。