title: Expression profiling and validation of potential reference genes during Paralichthys olivaceus embryogenesis.
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, alpha-Tub, EF1-alpha, 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.
authors.affiliation: Laboratory 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.
authors.forename: Qiwang
authors.initials: Q
authors.lastname: Zhong
authors.forename: Quanqi
authors.initials: Q
authors.lastname: Zhang
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Qi
authors.forename: Yanjie
authors.initials: Y
authors.lastname: Chen
authors.forename: Shuo
authors.initials: S
authors.lastname: Li
authors.forename: Yeying
authors.initials: Y
authors.lastname: Sun
authors.forename: Chunmei
authors.initials: C
authors.lastname: Li
authors.forename: Xun
authors.initials: X
authors.lastname: Lan
mesh_terms: D000818:Animals; D005412:Flatfishes / Q000196:embryology* / Q000235:genetics*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D012333:RNA, Messenger