title: Reference gene validation for quantification of gene expression during final oocyte maturation induced by diethylstilbestrol and di-(2-ethylhexyl)-phthalate in common carp.
abstract: Final oocyte maturation is the key step to successful spawning and fertilization. Quantitative real-time PCR (qPCR) is the technique of election to quantify the abundance of functional genes in such study. Reference gene is essential for correct interpretation of qPCR data. However, an ideal universal reference gene that is stable under all experimental circumstances has not been described. Researchers should validate their reference genes while performing qPCR analysis. The expression of 6 candidate reference genes: 18s rRNA, 28s rRNA, Cathepsin Z, Elongation factor 1-α, Glyceraldehyde-3-phosphate dehydrogenase and β-actin were investigated during final oocyte maturation induced by different compounds (DES and DEHP) in common carp (Cyprinus carpio). Four softwares (Bestkeeper, geNorm, NormFinder and RefFinder) were used to screen the most stable gene in order to evaluate their expression stability. The results revealed that EF1α was highly stable expressed when final oocyte maturation was induced by DES, while gapdh was the most stable gene when final oocyte maturation was induced by DEHP. Stable expressed reference gene selection is critical for all qPCR analysis to get accurate target gene mRNA expression information.
authors: Fisheries College, Jimei University, Xiamen 361021, China.
authors: Yanyan
authors: Y
authors: Shi
authors: School of Physics and Mechanical & Electrical Engineering, Xiamen University, Xiamen 361005, China.
authors: Jie
authors: J
authors: Lu
authors: Fisheries College, Jimei University, Xiamen 361021, China.
authors: Yilei
authors: Y
authors: Wang
authors: Fisheries College, Jimei University, Xiamen 361021, China. Electronic address: shwang@jmu.edu.cn.
authors: Shuhong
authors: S
authors: Wang
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D002347:Carps / Q000502:physiology*; D056663:Cathepsin Z / Q000235:genetics; D004051:Diethylhexyl Phthalate / Q000633:toxicity*; D004054:Diethylstilbestrol / Q000633:toxicity*; D015870:Gene Expression; D020869:Gene Expression Profiling; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D012337:RNA, Ribosomal, 18S; D060888:Real-Time Polymerase Chain Reaction; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D000199:Actins; D020648:Peptide Elongation Factor 1; D012337:RNA, Ribosomal, 18S; D014874:Water Pollutants, Chemical; D004054:Diethylstilbestrol; D004051:Diethylhexyl Phthalate; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D056663:Cathepsin Z
keywords: Cyprinus carpio; Di-(2-ethylhexyl)-phthalate; Diethylstilbestrol; Final oocyte maturation; Reference gene