利用荧光定量聚合酶链反应检测小鼠纹状体中Per1基因的表达规律

Expression Profiling of Per1 Gene in Mouse Striatum Detected Using Real Time RT-PCR

目的建立定量检测Per1 mRNA表达水平的系统,并检测小鼠纹状体中该基因的表达规律。方法提取小鼠纹状体总RNA,用Per1基因特异性引物进行聚合酶链反应(PCR),以荧光染料SYBR green I法进行实时检测。结果确定了Per1基因荧光定量的扩增和检测参数。证明在此条件下无非特异扩增,并且扩增效率在90%以上。利用此检测体系发现小鼠纹状体内Per1基因的表达呈节律性波动。结论所建立的方法能够定量测定Per1 mRNA表达水平,具有灵敏度高,线性范围广的特点。纹状体中Per1基因表达的节律性波动提示黑质纹状体系统间的相互调节可能具有昼夜节律性差异。

Objective To establish a method for quantifying the expression level of Perl in mouse striatum. Methods Total RNA isolated from mouse striatum was subjected to reverse transcription and amplification with Perl specific primers. SYBR green I was used for real-time PCR detection. Results Parameters in amplification and detection were determined. The quantification system was highly specific and the amplification efficiency was more than 90%. Expression of Perl showed a daily oscillation. Conclusion This system could used for quantifying mRNA level of Perl with high sensitivity and broad linearity. The time-dependent expression of Perl in striatum suggested that the interaction between Substantial Nigra and striatum may oscillate in a daily rhythmic base.