Characterization of reference genes for qRT-PCR normalization in rice-field eel (Monopterus albus) to assess differences in embryonic developmental stages, the early development of immune organs, and cells infected with rhabdovirus
文献类型： 外文期刊
作者： Liu, Yuchen 1 ; Wu, Shuwang 2 ; Jiang, Nan 2 ; Liu, Wenzhi 2 ; Zhou, Yong 2 ; Zeng, Lingbing 2 ; Zhong, Qiwang 3 ; Li, Zhong 2 ; Fan, Yuding 1 ;
作者机构： 1.Shanghai Ocean Univ, Natl Demonstrat Ctr Expt Fisheries Sci Educ, Shanghai 201306, Peoples R China
2.Chinese Acad Fishery Sci, Yangtze River Fisheries Res Inst, Wuhan 430223, Peoples R China
3.Jiangxi Agr Univ, Coll Biol Sci & Engn, Nanchang 330045, Jiangxi, Peoples R China
关键词： Rice-field eel; Monopterus albus; Reference gene; qRT-PCR; Developmental stage; Immune tissue; Cell culture
期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.622； 五年影响因子：4.799 ）
ISSN： 1050-4648
年卷期： 2022 年 120 卷
页码：
收录情况： SCI
摘要： Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) has become a popular technique to assess gene expression. Suitable reference genes are normally identified first to ensure accurate normalization. The aim of the present study was to select the most stable genes in embryonic developmental stages, the early development of immune organs, and cells infected with Chinese rice-field eel rhabdovirus (CrERV) of the rice-field eel (Monopterus albus). Four reference genes, including those encoding 18S ribosomal RNA (18SrRNA), beta actin (beta-actin), elongation factor 1 alpha (EF1alpha), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were assessed using geNorm, NormFinder, BestKeeper, and RefFinder software. Analyses indicated the stability ranking was 18SrRNA > beta-actin > GAPDH > EF1 alpha in the embryonic stage, with 18SrRNA as the most stable reference gene. For immunity-related organs at different developmental stages, the order in the thymus was beta-actin > GAPDH > EF1 alpha > 18SrRNA, with beta-actin as the most stable gene. In both spleen and kidney tissues, the rank order was EF1alpha > GAPDH > beta-actin > 18SrRNA, with EF1 alpha as the most stable gene. Furthermore, in rice-field eel kidney (CrE-K) cells infected with CrERV, the ranking was EF1alpha > beta-actin > GAPDH > 18SrRNA, with EF1 alpha as the most stable gene. The results for cells infected with CrERV were verified by testing signaling pathway genes catenin beta 1 (CTNNB1) and NOTCH1 based on the above four genes after virus infection in CrEK cells. This study laid the foundation for choosing suitable reference genes for immunity-related gene expression analysis in rice-field eel.
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