黄精实时荧光定量PCR内参基因的筛选

发布时间：2018-05-24 14:28

本文选题：黄精 + 实时荧光定量PCR　；参考：《基因组学与应用生物学》2017年11期

【摘要】：本研究采用实时荧光定量PCR(quantitative Real-t Time PCR,q RT-PCR)筛选适用于黄精(Polygonatum sibiricum)的内参基因。利用黄精转录组数据库,筛选8个候选内参基因,18S核糖体r RNA基因(18S-r RNA,18S)、肌动蛋白基因(Actin,ACT)、细胞色素基因(cytochrome,CYP)、3-磷酸甘油醛脱氢酶基因(GAPDH)、α-微管蛋白基因(α-Tubulin,TUA)、β-微管蛋白基因(β-Tubulin,TUB)、多聚泛素酶基因(ubiquitin 5,UBQ 5)和(ubiquitin 10,UBQ 10)。应用q RT-PCR技术检测这8个候选内参基因在黄精不同组织器官中(根,根茎,茎,叶,花和种子)的表达情况。利用Ge Norm、Norm Finder和Best Keeper 3种统计学软件综合评价8个内参基因的表达稳定性。研究结果表明,TUB在黄精不同组织部位中表达稳定性最好,运用q RT-PCR技术研究黄精器官基因表达时,可选用TUB作为内参基因。确定黄精q RT-PCR分析的合适内参基因,为后续相关基因表达研究奠定基础。
[Abstract]:In this study, real-time fluorescence quantitative PCR(quantitative Real-t Time PCR Q RT-PCR was used to screen the internal reference gene suitable for Polygonatum sibiricum. Using the Polygonatum transcriptome database, Eight candidate internal reference genes, including 18s ribosomal r RNA gene and 18S-r RNAs, actin gene (actin), cytochrome gene (cytochrome), cytochrome gene CYPN3-phosphoglyceraldehyde dehydrogenase gene, 伪 -TubulinTUAA, 尾 -TubulinTUBU, ubiquitin 5UBQ5 and ubiquitin 10UBQ5UBQ5, were screened. Q RT-PCR technique was used to detect the expression of the eight candidate internal reference genes in different tissues and organs (root, rhizome, leaf, flower and seed) of Rhizoma Polygonatum. Three statistical softwares, GE Norm Finder and Best Keeper, were used to evaluate the expression stability of 8 internal reference genes. The results showed that the expression stability of TUB was the best in different tissues of Polygonatum chinensis. When using Q RT-PCR technique to study the gene expression in the organ of semen flavescentis, TUB could be used as the internal reference gene. To determine the appropriate internal reference gene for Q RT-PCR analysis of Rhizoma Polygonatum, and to lay a foundation for further study on the expression of related genes.
【作者单位】：西北濒危药材资源开发国家工程实验室药用植物资源与天然药物化学教育部重点实验室陕西师范大学生命科学学院;
【基金】：国家自然科学基金(32170338) 陕西师范大学研究生创新基金项目《黄精种质资源研究》(2015CXB010) 陕西省科技厅科技计划项目《黄精优良种质资源选育及林下野生抚育技术研究》(2015SF309)共同资助
【分类号】：Q943.2;S567.239

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