caa a22 4500 606230084 CHVBK 20210128101212.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10695-015-0039-8 doi (NATIONALLICENCE)springer-10.1007/s10695-015-0039-8 Evaluation of potential internal references for quantitative real-time RT-PCR normalization of gene expression in red drum ( Sciaenops ocellatus ) [Elektronische Daten] [Bo-guang Sun, Yong-hua Hu] Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) has been used extensively for studying gene expression in diverse organisms including fish. In this study, with an aim to identify reliable reference genes for qRT-PCR in red drum (Sciaenops ocellatus), an economic fish species, we determined the expression stability of seven housekeeping genes in healthy and bacterium-infected red drum. Each of the selected candidate genes was amplified by qRT-PCR from the brain, gill, heart, intestine, kidney, liver, muscle, and spleen of red drum infected with or without a bacterial pathogen for 12 and 48h. The mRNA levels of the genes were analyzed with the geNorm and NormFinder algorithms. The results showed that in the absence of bacterial infection, translation initiation factor 3, NADH dehydrogenase 1, and QM-like protein may be used together as internal references across the eight examined tissues. Bacterial infection caused variations in the rankings of the most stable genes in a tissue-dependent manner. For all tissues, two genes sufficed for reliable normalization at both 12 and 48h post-infection. However, the optimal gene pairs differed among tissues and, for four of the examined eight tissues, between infection points. These results indicate that when studying gene expression in red drum under conditions of bacterial infection, the optimal reference genes should be selected on the basis of tissue type and, for accurate normalization, infection stage. Springer Science+Business Media Dordrecht, 2015 Housekeeping gene nationallicence Expression stability nationallicence Reference gene nationallicence Sciaenops ocellatus nationallicence Sun Bo-guang Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, 266071, Qingdao, People's Republic of China aut Hu Yong-hua Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, 266071, Qingdao, People's Republic of China aut Fish Physiology and Biochemistry Springer Netherlands 41/3(2015-06-01), 695-704 0920-1742 41:3<695 2015 41 10695 https://doi.org/10.1007/s10695-015-0039-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10695-015-0039-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sun Bo-guang Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, 266071, Qingdao, People's Republic of China aut NATIONALLICENCE 700 1- Hu Yong-hua Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, 266071, Qingdao, People's Republic of China aut NATIONALLICENCE 773 0- Fish Physiology and Biochemistry Springer Netherlands 41/3(2015-06-01), 695-704 0920-1742 41:3<695 2015 41 10695