Abstract
Differential expression of genes is crucial to embryogenesis. The analysis of gene expression requires appropriate references that should be minimally regulated during the embryonic development. To select the most stable genes for gene normalization, the expression profiles of eight commonly used reference genes (ACTB, GAPDH, rpL17, α-Tub, EF1-α, UbcE, B2M, and 18S rRNA) were examined during Japanese flounder (Paralichthys olivaceus) embryonic development using quantitative real-time polymerase chain reaction. It was found that all seven mRNA genes appeared to be developmentally regulated and exhibited significant variation of expression. However, further analyses revealed the stage-specific expression stability. Hence when normalization using these mRNA genes, the differential and stage-related expression should be considered. 18S rRNA gene, on the other hand, showed the most stable expression and could be recommended as a suitable reference gene during all embryonic developmental stages in P. olivaceus. In summary, our results provided not only the appropriate reference gene for embryonic development research in P. olivaceus, but also possible guidance to reference gene selection for embryonic gene expression analyses in other fish species.
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Acknowledgments
We thank Dr. Yuanmin Wu, from Funglyn BioTech Co., Ltd., for technical assistance. This work was supported by grants from the National High Technology Research and Development Program of China (No. 2006AA10A404) and the National Natural Science Foundation of China (No. 30671624).
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Zhong, Q., Zhang, Q., Wang, Z. et al. Expression Profiling and Validation of Potential Reference Genes During Paralichthys olivaceus Embryogenesis. Mar Biotechnol 10, 310–318 (2008). https://doi.org/10.1007/s10126-007-9064-7
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DOI: https://doi.org/10.1007/s10126-007-9064-7