Back to Search
Start Over
Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus)
- Source :
- Scientific Reports, Vol 10, Iss 1, Pp 1-10 (2020), Scientific Reports
- Publication Year :
- 2020
- Publisher :
- Nature Publishing Group, 2020.
-
Abstract
- Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a powerful and sensitive method used in gene expression analysis. Suitable reference genes, which are stable under all experimental circumstances and tissues significantly improve the accuracy of qRT-PCR data. In this study, the stability of six genes, namely, 18S ribosomal RNA (18s), beta-actin (actb), elongation factor 1-alpha (ef1α), glyceraldehyde-3-phosphate-dehydrogenase (gapdh), cathepsin D (ctsd), and beta-2-microglobulin (b2m) were evaluated as potential references for qRT-PCR analysis. The genes were examined in the hypothalamus-pituitary-ovary-liver (HPOL) axis throughout turbot ovarian development via using the geNorm, NormFinder and BestKeeper algorithms. Results showed that the most stable reference genes were ef1α, actb, and ctsd in the hypothalamus, pituitary, ovary and liver, respectively. The best-suited gene combinations for normalization were 18s, ef1α, and ctsd in the hypothalamus; actb, ctsd, and 18s in the pituitary; actb, and ctsd in the ovary; gapdh and ctsd in the liver. Moreover, the expression profile of estrogen receptor α (erα) manifested no significant difference normalization to the aforementioned best-suited gene during turbot ovarian development. However, no single gene or pair of genes is suitable as an internal control and account for the amplification differences among the four tissues during ovarian development. In summary, these results provide a basic data for the optimal reference gene selection and obtain highly accurate normalization of qRT-PCR data in HPOL axis-related gene expression analysis during turbot ovarian development.
- Subjects :
- 0301 basic medicine
Molecular biology
Gene Expression
Reproductive biology
Estrogen receptor
Cathepsin D
lcsh:Medicine
Ovary
Article
03 medical and health sciences
Peptide Elongation Factor 1
Reference genes
Gene expression
RNA, Ribosomal, 18S
medicine
Animals
lcsh:Science
Gene
Regulation of gene expression
Multidisciplinary
biology
Reverse Transcriptase Polymerase Chain Reaction
Gene Expression Profiling
lcsh:R
Gene Expression Regulation, Developmental
04 agricultural and veterinary sciences
biology.organism_classification
Actins
Turbot
030104 developmental biology
medicine.anatomical_structure
Flatfishes
040102 fisheries
0401 agriculture, forestry, and fisheries
Female
lcsh:Q
Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)
beta 2-Microglobulin
Subjects
Details
- Language :
- English
- ISSN :
- 20452322
- Volume :
- 10
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....9c1ccb8ff9c6ee344ddb21b1919c0004