葡萄实时定量PCR中稳定内参基因的筛选

发布时间：2018-05-16 04:35

本文选题：葡萄 + ge　；参考：《果树学报》2016年03期

【摘要】：【目的】通过ge Norm软件筛选实时荧光定量PCR稳定内参基因,基于多内参基因评价体系进行目的基因相对表达量的计算。【方法】获得不同葡萄品种叶片和果皮中6个候选内参基因的循环阈值(Ct),利用ge Norm软件计算内参基因平均表达稳定性数值M和基因配对差异值V,从而判断内参基因最适组合。【结果】发现候选内参基因表达稳定性由高到低的排列顺序为Vv EF1r=Vv EF1-αVv GAPDHVv ACTINVv ACT1Vv UBQ,不同组织分别分析均发现Vv EF1-α和Vv EF1r的稳定性最高,且基因配对差异值V2/3为0.104,所以内参基因的最适组合为Vv EF1-α和Vv EF1r。利用ge Norm软件计算得到的多内参基因评价体系的标准化因子可应用于目的基因的相对表达量分析。【结论】ge Norm软件筛选实时荧光定量PCR稳定内参基因的方法可以应用到多条件和多组织的不同植物样本中,并用于目的基因表达谱的研究,具有重要的广泛应用价值。
[Abstract]:[objective] to screen the stable internal reference gene of real-time fluorescent quantitative PCR by ge Norm software. The relative expression of target gene was calculated based on the multiple internal reference gene evaluation system. [methods] the cyclic threshold of 6 candidate internal reference genes in leaves and pericarp of different grape varieties was obtained, and the GE Norm software was used to calculate the internal reference gene. The average expression stability value M and gene pairing difference value V were used to judge the optimal combination of endogenous reference genes. [results] it was found that the sequence of stability of candidate internal reference genes from high to low was VV EF1r=Vv EF1- 伪 Vv GAPDHVv ACTINVv ACT1Vv UBQs, and different tissues. The stability of Vv EF1- 伪 and Vv EF1r were found to be the highest. The difference of gene pairing value V2 / 3 is 0.104, so the optimal combination of internal reference gene is VvEF1- 伪 and VvEF1r. The standardized factors calculated by ge Norm software can be used to analyze the relative expression of the target gene. [conclusion] the method of screening real-time fluorescent quantitative PCR stable endogenous reference gene by ge Norm software can be used. To apply to different plant samples with multiple conditions and multiple tissues, And used in the study of target gene expression profile, has an important and extensive application value.
【作者单位】：上海农业科学院林木果树研究所;上海交通大学农学与生物技术学院;
【基金】：国家葡萄产业技术体系项目(CARS-30-9) 上海市农委科技兴农推广项目[沪农科推字(2012)第1-2号]
【分类号】：S663.1

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