Evaluation of potential reference genes for real time RT-PCR studies in Atlantic halibut ( Hippoglossus Hippoglossus L.); during development, in tissues of healthy and NNV-injected fish, and in anterior kidney leucocytes

摘要: Background: Real time RT-PCR has become an important tool for analyzing gene expression in fish. Although several housekeeping genes have been evaluated Atlantic halibut (Hippoglossus Hippoglossus L.), appropriate reference low copy mRNA transcripts at the earliest developmental stages not identified. No attempts reported to identify suitable infected with NNV or stimulated leucocytes. In this study, β-actin1 (ACTB1), elongation factor 1 alpha (EF1A1), hypoxanthine-guanine phosphoribosyltransferase (HPRT1), ribosomal protein L7 (RPL7), tubulin beta 2C (Tubb2C), and ubiquitin-conjugating enzyme (UbcE) were as normalization of real data during development, tissue healthy NNV-infected fish, vivo vitro anterior kidney Results: The all six was relatively stable from unfertilized egg until 12 day degrees post fertilization (ddpf ). However, none selected found be stably expressed throughout development. levels increased 18 ddpf, when zygotic transcription is likely activated, stabilized different points. Excel-based software programs BestKeeper, geNorm, NormFinder ranked EF1A1 UbcE best candidate before activation transcription, RPL7 candidates after hatching. also listed exploring various organs, both non-injected fish mockand NNV-injected None optimal Conclusion: Generally, it that showed least variation, HPRT1 intermediate genes, ACTB1 Tubb2C ones. can recommended ConA-PMA a good other experimental setups. This study emphasizes need evaluation, universal