Selection of Reference Genes in Transcription Analysis of Gene Expression of the Mandarin Fish, Siniperca chuasti.
Selection of Reference Genes in Transcription Analysis of Gene Expression of the Mandarin Fish, Siniperca chuasti.
复制标题
DOI：
10.3724/sp.j.1141.2010.02141
复制
发表时间：
2010-04
期刊：
影响因子：
--
通讯作者：
Ruixue Zhou;Tao Meng;Haining Meng;D. Cheng;Shiyu Bin;Jia Cheng;G. Fu;W. Chu;Jianshe Zhang-
中科院分区：
文献类型：
--
作者：
Ruixue Zhou;Tao Meng;Haining Meng;D. Cheng;Shiyu Bin;Jia Cheng;G. Fu;W. Chu;Jianshe Zhang-
目前，基因表达的转录分析通常使用管家基因作为标准化的对照。采用实时荧光定量PCR（qPCR）技术，分析了鳜鱼GAPDH、beta-actin和18 SrRNA 3个管家基因在6种组织和5个发育阶段的表达水平。使用geNorm程序分析表达水平的差异。结果表明，β-actin是发育阶段最稳定的基因，GAPDH在不同组织中最稳定。而18 S rRNA在发育过程中的表达是差异调节的，这表明它适合作为发育水平基因表达正常化的内部对照。总的来说，数据表明，两个最稳定的管家基因足以准确校准S。恰齐。本研究为常规PCR或qPCR技术在基因表达分析中管家基因的选择和标准化提供了有说服力的参考和方法学依据。
At present, transcription analysis of gene expression commonly uses housekeeping genes as control for normalization. In this study, the expression levels of three housekeeping genes including GAPDH, beta-actin, and 18S rRNA in six tissues and five developmental stages of the Mandarin fish Siniperca chuatsi were assayed with quantitative real-time PCR (qPCR). Differences in expression levels were analyzed using geNorm program. The results demonstrate that beta-actin is the most stable gene at developmental stages and GAPDH is the most stable in different tissues. While 18S rRNA expression during development is differentially regulated, which indicates it is suitable as an internal control for gene expression normalization at the developmental level. Overall, the data suggest that the two most stable housekeeping genes are enough to accurately calibrate gene expression in S. chuatsi. The significance of this study provided convincing references and methodology for housekeeping gene selection and normalization in gene expression analysis with regular PCR or qPCR.