【摘要】 为应用实时荧光定量PCR(qPCR)技术分析革胡子鲶主要组织相容性复合体(major histocompatibility complex,MHC)基因的表达谱,本研究利用Primer 5软件设计扩增革胡子鲶MHC基因片段的引物并进行qPCR评估。结果显示:引物MHCF1-MHCR3在qPCR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99.3%,R2值为0.998。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。本研究为革胡子鲶MHCⅠ基因表达的qPCR分析奠定了基础。更多还原

【Abstract】 In order to analyze the MHC gene expression profiles in Clarias gariepinus by using real-time quantitative PCR(q PCR)technique, primer pairs for amplifying the MHC gene fragments of C. gariepinus were designed using Primer 5.0 software and then assessed by means of general PCR and q PCR amplification, respectively. The results of q PCR evaluation showed that melt curve of the primer pair named MHCF1-MHCR3 presented a single sharp peak, and standard curve displayed that its amplification efficiency was 99.3% and R2 value was 0.998. The results indicated that the primer pair had good specificity and efficiency of amplification as well as met the criteria of q PCR amplification. This work laid a foundation for analyzing MHC gene expression of C. gariepinus by using q PCR technique.更多还原