Quantitative real time RT-PCR study of pathogen-induced gene expression in rock bream (Oplegnathus fasciatus): Internal controls for data normalization

摘要: Rock bream Oplegnathus fasciatus is an important economic fish species. In this study, we evaluated the appropriateness of six housekeeping genes as internal controls for quantitative real-time PCR (RT-qPCR) analysis gene expression in rock before and after pathogen infection. The selected eight tissues infected with Vibrio alginolyticus or megalocytivirus was determined by RT-qPCR, data were analyzed geNorm NormFinder algorithms. results showed that infection, mediator RNA polymerase II transcription subunit 8 β-actin ranked most stable across examined tissues. After bacterial viral stabilities varied to significant extents tissue-dependent manners, no single pair identified suitable references all either stimuli. addition, majority tissues, during infection differed from those Nevertheless, optimum reference each tissue under different conditions. Taken together, these indicate type nature infectious agent used study can influence choice normalization factors, will provide a useful guidance selection future RT-PCR bream.