鹅GHR基因荧光定量PCR检测方法的建立

Establishment of TaqMan Real-time PCR for Detection Goose GHR Gene

采用PCR方法克隆了鹅生长激素受体(GHR)基因,构建了重组质粒pMD-18T-GHR,并将其作为标准阳性模板。通过对反应条件进行优化,以定量的10倍系列稀释的质粒pMD-18T-GHR为标准品进行荧光定量PCR扩增,建立了检测GHR的荧光定量PCR方法。结果显示,该方法构建的标准曲线线性关系良好,建立的GHR基因荧光定量PCR检测方法灵敏度高、特异性强(可以检测出低于10个拷贝/μL的样品),准确可靠。籽鹅和莱茵鹅GHRmRNA出壳到90日龄生长过程中肝脏中的表达量不同,30日龄不同组织表达量各有变化特点。

Growth hormone receptor （GHR）of goose was cloned by PCR,and the recombinant,designated pMD-18T-GHR was constructed. In order to establish the fluorescent quantitative PCR（FQ-PCR）method that can detect the GHR, we have optimized the reaction conditions,amplified by FQ-PCR using serial 10-fold dilution of the pMD-18T-GHR as standard preparation. The results showed that the standard curve possess a good linear relationship by the method, construct the high sensitive and specific （can be detected in less than 10 copies/μL sample）FQ-PCR method,accurate and reliable. The expression of GHR mRNA was different in liver from birth to 90-day-age,and there was different changing pattern in other tissues at 30-day-old in Zi goose and Rhine.