肉雏鸡肝细胞谷胱甘肽过氧化物酶mRNA表达产物的RT-PCR检测

Quantitative analysis for cellular glutathione peroxidase mRNA in broiler hepar

细胞谷胱甘肽过氧化物酶 (cGPX)是动物体内一种重要的抗氧化酶。建立了利用RT PCR并以 β 肌动蛋白 (β Actin)为内参照定量分析cGPXmRNA表达的方法。提取肉鸡肝总RNA ,经反转录和PCR扩增 ,PCR产物的电泳条带于VDS摄像系统扫描灰度。寻求PCR线性扩增范围 ,确定RT PCR的最佳循环次数和模板量。通过计算cGPXPCR产物与 β 肌动蛋白PCR产物的灰度之比 ,即可得出cGPXmRNA的相对含量。

Cellular glutathione peroxidase(cGPX) is an important antioxidase in animals. This experiment setsup a method that can beused for quantitative analysis cGPX mRNA in RT PCR process. β Actin is used forinternal standard. Total RNA was extracted from livertissue of broiler. The mRNA was reverse transcripted and the cDNA fragmentsof cGPX gene and β Actin gene were amplified by RT PCR using the gene specific primers.The integrated optical density(IOD) was showed by scanning the electrophoresis bands of PCR products with VDS system. By studying the range of PCR products linearity amplification, the best of all cycle number and the volume of template(cDNA) were made certain. The relative concentration of cGPX mRNA can be calculated by the IOD ratio of cGPX and β Actin.