实时荧光定量PCR检测小鼠脑组织中3-羟基丁酸受体基因PUMA-G的转录表达

[摘要] 目的 检测小鼠海马、下丘脑、额叶皮质和颞叶皮质组织中的3-羟基丁酸和烟酸受体基因PUMA-G（protein upregulated in macrophages by IFN-g）的转录表达。方法 采用试剂盒法提取脑组织总RNA,分光光度法测定RNA的含量和纯度,变性凝胶电泳检测RNA的完整性,实时荧光定量PCR技术检测PUMA-G的转录表达。结果 与阳性对照组（心肌组织）一样,PUMA-G在小鼠海马、下丘脑、额叶皮质和颞叶皮质组织中均有转录表达,其循环阈值（cycle threshold, Ct）低于40个循环数。但是各种脑组织中的转录表达水平不同,其中下丘脑的表达量最高与阳性对照组（35.15±0.31）相似,其Ct值为35.39±0.57,而颞叶皮质中的表达最低,其Ct值为38.67±0.90,海马和额叶皮质的Ct值分别为37.59±0.60和37.75±0.51。结论 通过实时荧光定量PCR技术检测到PUMA-G在小鼠脑组织中的转录表达。

Detection of PUMA-G transcription in mice brain tissues by Real-time fluorescence quantitative PCR

Objective To demonstrated that the receptor for 3-hydroxyburate (3-HB) and Nicotinic Acid, namely, protein upregulated in macrophages by IFN-g (PUMA-G), was expressed in mice brain tissues, including Hippocampus, Subthalamic, Frontal cortex and Temporal cortex. Methods Extract whole RNA from mice brain tissues by kits, the concentration and purity of RNA were detected by spectroscopic analysis, the integrality of RNA was observed by denaturing gel electrophoresis, the transcript expression of PUMA-G was detected by Real-time fluorescence quantitative PCR. Results In all mouse brain tissues and positive group (heart tissue), the cycle threshold less than 40, showed that the transcript expression of PUMA-G was detected. But there were different transcript expression level in different brain tissues, the highest in the Subthalamic tissues (Ct: 35.39±0.57) as the positive group (35.15±0.31) and the lowest in the Frontal cortex tissues (Ct: 38.67±0.90). In Hippocampus and Temporal cortex tissues, the Ct value was 37.59±0.60 and 37.75±0.51 respectively. Conclusion Here described the transcript expression of PUMA-G in mice brain tissues by Real-time fluorescence quantitative PCR.