血鹦鹉TYR基因表达qPCR分析的引物设计与评估

Design and Evaluation of Primers for Analysis of TYR Gene Expression in Blood Parrot(Amphilophus)by Using Real-Time Fluorescence Quantitative PCR Technique

为了解色素相关基因和体色之间的关系,应用实时荧光定量PCR(qPCR)技术分析血鹦鹉的酪氨酸酶(tyrosinase,TYR)基因的表达谱,通过Primer 5软件设计扩增血鹦鹉TYR基因片段的引物并进行qPCR评估研究。结果显示,引物TYR2F-TYR2R在q PCR扩增时,融解曲线为单一尖锐峰;标准曲线分析显示,引物的扩增效率为103.3%,R2值为0.994。上述结果说明该对引物具有良好的扩增特异性和扩增效率,满足了qPCR扩增的要求,该研究结果可为血鹦鹉TYR基因表达的qPCR分析奠定基础。

To understand the relationship between the related genes and body color pigment, the application of real-time fluorescent quantitative PCR （qPCR） technical analysis blood parrot tyrosinase, tyrosinase, TYR） gene expression profile, this research designed blood pan＇or TYR gene primers and qPCR assessment through the Primer amplification 5 software. The results showed that the melting curve was a single sharp peak when the primer TYR2F-TYR2R was amplified by qPCR. The standard curve analysis showed that the amplification efficiency of primers was 103.3% and R2 was 0.994. The above results showed that the primers have good amplification specificity and amplification efficiency and meet the requirement of qPCR amplification. This study will lay a foundation for the qPCR analysis of TYR gene expression.