本文關(guān)鍵詞：實(shí)時(shí)熒光定量PCR分析中毛果楊內(nèi)參基因的篩選和驗(yàn)證，由筆耕文化傳播整理發(fā)布。

實(shí)時(shí)熒光定量PCR分析中毛果楊內(nèi)參基因的篩選和驗(yàn)證

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]山西師范大學(xué)生命科學(xué)學(xué)院,臨汾041000; [2]中國(guó)醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥用植物研究所,北京100193

文章摘要：實(shí)時(shí)熒光定量PCR（qRT-PCR）技術(shù)具有高靈敏性、高保真性和高特異性，被廣泛應(yīng)用于基因表達(dá)的分析。在數(shù)據(jù)處理過程中，選用穩(wěn)定表達(dá)的基因作為內(nèi)參基因?qū)?zhǔn)確分析實(shí)驗(yàn)結(jié)果非常關(guān)鍵。以毛果楊（Populustrichocarpa）的不同組織以及鋅脅迫下的組培苗為材料，使用熒光定量PCR方法分析了TUA8、TUB6、ubfquitin、GAPDH、actin、18SrRNA和EF1α7個(gè)看家基因的表達(dá)情況。通過geNorm、NormFinder和BestKeeper3個(gè)程序的綜合分析，發(fā)現(xiàn)actin、u6iquitin、EF1α和18SrRNA的穩(wěn)定性較好�？捎米髅麠罨虮磉_(dá)研究的內(nèi)參基因：而TUB6在不同組織中穩(wěn)定性最差：GAPDH在鋅脅迫下的組織中穩(wěn)定性最差，因此不適宜作為內(nèi)參基因。毛果楊NAC基因的表達(dá)分析，進(jìn)一步驗(yàn)證了上述結(jié)果。該研究對(duì)采用qRT-PCR方法分析毛果楊基因表達(dá)過程中內(nèi)參基因的選擇具有指導(dǎo)作用，，同時(shí)對(duì)揭示NAC基因的功能也有一定的意義。

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.

文章關(guān)鍵詞：

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

課題項(xiàng)目：國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展規(guī)劃（No.2012CB14502）和國(guó)家自然科學(xué)基金（No.31070534）

作者信息：會(huì)員可見

  本文關(guān)鍵詞：實(shí)時(shí)熒光定量PCR分析中毛果楊內(nèi)參基因的篩選和驗(yàn)證，由筆耕文化傳播整理發(fā)布。