Selection of reference genes for expression studies with fish myogenic cell cultures.
Selection of reference genes for expression studies with fish myogenic cell cultures.
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DOI：
10.1186/1471-2199-10-80
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发表时间：
2009-08-10
影响因子：
--
通讯作者：
Johnston IA
中科院分区：
文献类型：
--
作者：
Bower NI;Johnston IA
相对较少的研究已经使用细胞培养系统来研究基因表达和鱼类肌肉发生的调节。为了从定量实时PCR产生稳健的数据，需要使用在所有实验样品中具有稳定表达的内参基因对mRNA水平进行归一化。我们已经研究了八个候选基因的表达，以确定合适的参考基因用于从大西洋鲑鱼（萨尔莫salar L.）的原代肌细胞培养。使用软件分析包geNorm、Normfinder和Best keeper根据基因在肌源性分化过程中在42个样品中的稳定性对基因进行排序。初步结果显示，几个候选基因在整个生肌培养中表现出稳定的表达，而Sdha被鉴定为最不稳定的基因。用geNorm、Normfinder和Bestkeeper进一步分析，发现Ef1α、Hprt1、Ppia和RNApolII稳定表达。与从这些基因中的任何三个的组合获得的几何平均值归一化的数据的比较显示没有显著差异，表明这些基因的任何组合都是有效的。Hprt1、Ef1α、Ppia和RNApolII中任意三个的几何平均值适用于大西洋鲑鱼原代生肌培养物中基因表达数据的标准化。
Relatively few studies have used cell culture systems to investigate gene expression and the regulation of myogenesis in fish. To produce robust data from quantitative real-time PCR mRNA levels need to be normalised using internal reference genes which have stable expression across all experimental samples. We have investigated the expression of eight candidate genes to identify suitable reference genes for use in primary myogenic cell cultures from Atlantic salmon (Salmo salar L.). The software analysis packages geNorm, Normfinder and Best keeper were used to rank genes according to their stability across 42 samples during the course of myogenic differentiation. Initial results showed several of the candidate genes exhibited stable expression throughout myogenic culture while Sdha was identified as the least stable gene. Further analysis with geNorm, Normfinder and Bestkeeper identified Ef1α, Hprt1, Ppia and RNApolII as stably expressed. Comparison of data normalised with the geometric average obtained from combinations of any three of these genes showed no significant differences, indicating that any combination of these genes is valid. The geometric average of any three of Hprt1, Ef1α, Ppia and RNApolII is suitable for normalisation of gene expression data in primary myogenic cultures from Atlantic salmon.
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影响因子：
--
作者：
Silver N;Best S;Jiang J;Thein SL
通讯作者：
Thein SL
影响因子：
4.3
作者：
Deindl, E;Boengler, K;Schaper, W
通讯作者：
Schaper, W
影响因子：
3.6
作者：
KOUMANS, JTM;AKSTER, HA;OSSE, JWM
通讯作者：
OSSE, JWM
影响因子：
2.9
作者：
Solanas, M;Moral, R;Escrich, E
通讯作者：
Escrich, E
DOI：
10.1016/j.bbrc.2008.01.006
发表时间：
2008-03-14
影响因子：
3.1
作者：
Nishimura, Masuhiro;Nikawa, Takeshi;Ikeda, Muneharu
通讯作者：
Ikeda, Muneharu