实时荧光定量PCR分析中毛果杨内参基因的筛选和验证

  本文关键词：实时荧光定量PCR分析中毛果杨内参基因的筛选和验证，由笔耕文化传播整理发布。

实时荧光定量PCR分析中毛果杨内参基因的筛选和验证

Selection and Validation of Reference Genes for Quantitative RT-PCR Analysis of Gene Expression in Populus trichocarpa

[1] [2] [3] [4] [5]

Xiaojuan Su, Baoguo Fan, Lichai Yuan, Xiuna Cui, Shanfa Lu（1 College of Life Sciences, shanxi Normal University, Linfen 041000, China; 2Institute of Medicinal Plant Development

[1]山西师范大学生命科学学院,临汾041000; [2]中国医学科学院北京协和医学院药用植物研究所,北京100193

文章摘要：实时荧光定量PCR（qRT-PCR）技术具有高灵敏性、高保真性和高特异性，被广泛应用于基因表达的分析。在数据处理过程中，选用稳定表达的基因作为内参基因对准确分析实验结果非常关键。以毛果杨（Populustrichocarpa）的不同组织以及锌胁迫下的组培苗为材料，使用荧光定量PCR方法分析了TUA8、TUB6、ubfquitin、GAPDH、actin、18SrRNA和EF1α7个看家基因的表达情况。通过geNorm、NormFinder和BestKeeper3个程序的综合分析，发现actin、u6iquitin、EF1α和18SrRNA的稳定性较好。可用作毛果杨基因表达研究的内参基因：而TUB6在不同组织中稳定性最差：GAPDH在锌胁迫下的组织中稳定性最差，因此不适宜作为内参基因。毛果杨NAC基因的表达分析，进一步验证了上述结果。该研究对采用qRT-PCR方法分析毛果杨基因表达过程中内参基因的选择具有指导作用，，同时对揭示NAC基因的功能也有一定的意义。

Abstr：Quantitative RT-PCR （qRT-PCR） has been widely used in gene expression analysis because of its sensitivity, specificity, and reproducibility. Application of suitable reference genes to normalize qRT-PCR data is critical in analyzing PCR results. We analyzed the expression patterns of 7 housekeeping genes, including TUA8, TUB6, ubiquitin, GAPDH, actin, 18S rRNA and EFla, in various tissues of greenhouse-grown Populus trichocarpa and Zn-treated in vitro plantlets. The stability of housekeeping gene expression was analyzed with use of 3 software packages, including geNorm, Norm- Finder, and BestKeeper. The genes actin, ubiquitin, EFla and 18S rRNA were suitable reference genes for efficient normalization of qRT-PCR data, whereas TUB6 and GAPDH were not suitable for analysis of greenhouse-grown plants and Zn-treated plantlets, respectively. These findings were confirmed by comparative profiling of 4 P. trichocarpa NAC genes. This study provides useful information for reference gene selection in qRT-PCR analysis of gene expression in P. trichocarpa. It is also helpful to elucidate the function of P. trichocarpa NAC genes.

文章关键词：

Keyword：:NAC, Populus trichocarpa, qRT-PCR, reference gene, Zn stress

课题项目：国家重点基础研究发展规划（No.2012CB14502）和国家自然科学基金（No.31070534）

作者信息：会员可见

  本文关键词：实时荧光定量PCR分析中毛果杨内参基因的筛选和验证，由笔耕文化传播整理发布。