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Published on: October 27, 2017
Expression profiling and validation of potential reference genes during Paralichthys olivaceus embryogenesis
Qiwang Zhong1, Quanqi Zhang, Zhigang Wang
1Laboratory of Marine Genetics and Breeding, College of Marine Life Science, Ocean University of China, No.5 Yushan Road, Qingdao 266003, People's Republic of China.
Marine Biotechnology (New York, N.Y.)
|January 25, 2008
Summary
Selecting stable reference genes is vital for analyzing gene expression during Japanese flounder embryogenesis. The 18S rRNA gene demonstrated the most consistent expression, making it ideal for normalization across all developmental stages.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genomics
Background:
- Accurate gene expression analysis in embryogenesis relies on stably expressed reference genes for normalization.
- Identifying reliable reference genes is crucial for understanding developmental processes in species like the Japanese flounder (Paralichthys olivaceus).
Purpose of the Study:
- To evaluate the expression stability of commonly used reference genes during Japanese flounder embryonic development.
- To identify the most suitable reference gene for accurate gene expression normalization in this species.
Main Methods:
- Quantitative real-time polymerase chain reaction (qPCR) was employed to analyze the expression profiles of eight candidate reference genes.
- Expression levels were assessed across various embryonic developmental stages of Japanese flounder.
Main Results:
- Seven out of eight tested mRNA genes (ACTB, GAPDH, rpL17, alpha-Tub, EF1-alpha, UbcE, B2M) exhibited significant developmental regulation and expression variability.
- The 18S ribosomal RNA (rRNA) gene displayed the most stable expression throughout embryonic development.
- Stage-specific expression patterns were observed for the mRNA reference genes, necessitating careful consideration during normalization.
Conclusions:
- The 18S rRNA gene is recommended as a reliable reference gene for gene expression studies during Japanese flounder embryonic development.
- Researchers should consider the stage-specific expression of mRNA reference genes when performing normalization in P. olivaceus embryogenesis.
- This study provides valuable guidance for selecting appropriate reference genes in fish embryonic development research.
