猪IGF1、EGFR、EGF、β-actin基因荧光定量PCR标准曲线的建立

发布时间：2018-03-18 16:04

本文选题：猪　切入点：修复基因　出处：《河北农业大学学报》2017年05期 　论文类型：期刊论文

【摘要】：为建立用荧光定量PCR方法检测猪IGF1、EGFR、EGF、β-actin基因表达量的标准曲线,本试验根据目的基因在NCBI上CDS区序列,设计合成引物,构建目的基因重组质粒,采用SYBR GreenI荧光定量RT-PCR的检测方法,构建检测猪IGF1、EGFR、EGF、β-actin基因表达量的标准曲线。结果显示,由pEASY-T1目的基因所构建的标准曲线线性关系良好,线性回归系数(R2)均大于或等于0.99,起始模板与Ct值之间呈良好的线性关系,溶解曲线峰值单一,表明引物具有很好的特异性,结果准确可靠。本研究成功构建了用荧光定量RT-PCR方法检测猪IGF1、EGFR、EGF、β-actin基因表达量的标准曲线,为后续检测基因相对表达量的变化奠定基础。
[Abstract]:In order to establish a standard curve for detecting the expression of EGF1, 尾 -actin gene in porcine IGF1-EGFR, 尾 -actin gene, primers were designed and synthesized according to the sequence of CDS region of the target gene on NCBI, and the recombinant plasmid of the target gene was constructed. The detection method of SYBR GreenI fluorescence quantitative RT-PCR was used to detect the expression of EGF, 尾 -actin gene. A standard curve was constructed to detect the expression of EGF, 尾 -actin gene in pig IGF1 gene. The results showed that the standard curve constructed by pEASY-T1 gene had a good linear relationship, and the linear regression coefficient (R2) was greater than or equal to 0.99, and there was a good linear relationship between the initial template and Ct value. The peak value of the dissolution curve was single, which indicated that the primer had good specificity and the results were accurate and reliable. In this study, a standard curve for the detection of EGF1, 尾 -actin gene expression in pig IGF1 was successfully constructed by fluorescence quantitative RT-PCR. It lays a foundation for the subsequent detection of the relative expression of genes.
【作者单位】：河北农业大学兽医诊断中心/农业部动物疫病病原生物学华北科学观测实验站;衡水市农牧局;伊犁特克斯县畜牧局兽医站;
【基金】：国家自然基金项目(31470125)
【分类号】：Q78;S828

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