摘要
以革胡子鲇(Clarias gariepinus)为试验对象,对其生长激素(GH)基因进行荧光定量PCR(qPCR)的引物设计与评估。将合成的5对引物通过常规PCR电泳结果选择出符合条件的GHF-GHR引物,并进行qPCR引物的评估。结果显示:扩增GH基因片段的引物GHF-GHR扩增时熔解曲线为单一峰,说明引物具有良好的特异性;通过标准曲线得出GHF-GHR引物的扩增效率为93.3%,标准曲线的R2值为0.982,说明引物的扩增效率良好。上述结果说明GHF-GHR引物能用于待检样本的GH基因表达的qPCR分析。
This experiment focused on the design and evaluation of primers for growth hormone(GH)gene in Clarias gariepinus using fluorescence quantitative PCR(qPCR).Five pairs of primers were synthesized,and the GHF-GHR primer pair was selected based on the results of conventional PCR electrophoresis.Subsequently,the GHF-GHR primer pair was evaluated for qPCR.The results indicated that the amplification curve of the GH gene fragment with the GHF-GHR primer pair showed a single peak,demonstrating good specificity of the primers.The amplification efficiency of the GHF-GHR primer pair was determined to be 93.3%based on the standard curve,with an R 2 value of 0.982,indicating a high amplification efficiency of the primers.These results suggest that the GHF-GHR primer pair is suitable for qPCR analysis of GH gene expression in the test samples.
作者
蓝一
何净慧
王晓梅
王茜
LAN Yi;HE Jinghui;WANG Xiaomei;WANG Qian(College of Aquaculture,Tianjin Agricultural University,Tianjin 300384,China;Key Laboratory of Aquatic-Ecology and Aquaculture of Tianjin,Tianjin 300384,China)
基金
国家自然科学基金(31672264)
天津市淡水养殖产业体系创新团队建设项目(ITTFRSA2021000)。
