Evaluation of suitable reference genes for gene expression studies in bovine muscular tissue.
Evaluation of suitable reference genes for gene expression studies in bovine muscular tissue.
复制标题
评估合适的参考基因用于牛肌肉组织中的基因表达研究。
DOI：
10.1186/1471-2199-9-79
复制
发表时间：
2008-09-11
影响因子：
--
通讯作者：
Dunner S
中科院分区：
文献类型：
--
作者：
Pérez R;Tupac-Yupanqui I;Dunner S
实时逆转录酶定量聚合酶链反应（real-time RTqPCR）是一种用于测量mRNA物种拷贝数的技术，作为确定参与不同生物过程的关键基因的一种方法。然而，这些关键基因的表达水平可能在组织或细胞中变化，这不仅是由于差异表达的结果，而且还由于不同的因素，包括参考基因的选择以标准化靶基因的表达水平;因此参考基因的选择对于表达研究至关重要。为此，在牛肌肉组织中研究了10个候选参考基因。评估了三组中包括的十个候选参考基因的稳定性值：所谓的“经典管家”基因（18 S、GAPDH和ACTB）、在其他组织上进行的表达研究中使用的第二组基因（B2 M、RPII、UBC和HMBS）和第三组新基因（SF 3A 1、EEF 1A 2和CASC 3）。使用三种不同的统计算法通过geNorm、Normandy和Bestkeeper产生的稳定性度量对基因进行排名。这三种方法倾向于在最稳定表达的基因和肌肉组织中最少的基因上达成一致。EEF 1A 2和HMBS之后是SF 3A 1、ACTB和CASC 3可以被认为是稳定的参考基因，而B2 M、RPII、UBC和GAPDH将不合适。尽管rRNA-18 S稳定性指标似乎在可接受范围内，但不建议使用，因为其合成调节不代表mRNA水平。基于geNorm算法，我们提出使用SF 3A 1，EEF 1A 2和HMBS三个基因作为肌肉表达研究中实时RTqPCR标准化的参考。
Real-time reverse transcriptase quantitative polymerase chain reaction (real-time RTqPCR) is a technique used to measure mRNA species copy number as a way to determine key genes involved in different biological processes. However, the expression level of these key genes may vary among tissues or cells not only as a consequence of differential expression but also due to different factors, including choice of reference genes to normalize the expression levels of the target genes; thus the selection of reference genes is critical for expression studies. For this purpose, ten candidate reference genes were investigated in bovine muscular tissue. The value of stability of ten candidate reference genes included in three groups was estimated: the so called 'classical housekeeping' genes (18S, GAPDH and ACTB), a second set of genes used in expression studies conducted on other tissues (B2M, RPII, UBC and HMBS) and a third set of novel genes (SF3A1, EEF1A2 and CASC3). Three different statistical algorithms were used to rank the genes by their stability measures as produced by geNorm, NormFinder and Bestkeeper. The three methods tend to agree on the most stably expressed genes and the least in muscular tissue. EEF1A2 and HMBS followed by SF3A1, ACTB, and CASC3 can be considered as stable reference genes, and B2M, RPII, UBC and GAPDH would not be appropriate. Although the rRNA-18S stability measure seems to be within the range of acceptance, its use is not recommended because its synthesis regulation is not representative of mRNA levels. Based on geNorm algorithm, we propose the use of three genes SF3A1, EEF1A2 and HMBS as references for normalization of real-time RTqPCR in muscle expression studies.
登录
查看更多内容
影响因子：
14.9
作者：
Imbeaud S;Graudens E;Boulanger V;Barlet X;Zaborski P;Eveno E;Mueller O;Schroeder A;Auffray C
通讯作者：
Auffray C
影响因子：
3.7
作者：
de Jonge HJ;Fehrmann RS;de Bont ES;Hofstra RM;Gerbens F;Kamps WA;de Vries EG;van der Zee AG;te Meerman GJ;ter Elst A
通讯作者：
ter Elst A
影响因子：
3.5
作者：
Erkens T;Van Poucke M;Vandesompele J;Goossens K;Van Zeveren A;Peelman LJ
通讯作者：
Peelman LJ
影响因子：
2.7
作者：
Dheda, K;Huggett, JF;Zumla, A
通讯作者：
Zumla, A
影响因子：
4.8
作者：
Mahoney, DJ;Parise, G;Tarnopolsky, MA
通讯作者：
Tarnopolsky, MA