Validation of reference genes of grass carp Ctenopharyngodon idellus for the normalization of quantitative real-time PCR

• Published in: Biotechnology letters 2010-08, Vol.32 (8), p.1031-1038
• Main Authors: Ye, Xing, Zhang, Lili, Dong, Haiyan, Tian, Yuanyuan, Lao, Haihua, Bai, Junjie, Yu, Lingyun
• Format: Article
• Language: English
• Subjects: Animals; Applied Microbiology; Bacteria; Bacterial infection; Bacterial Infections - metabolism; Bacterial Infections - veterinary; Bacterial Toxins - genetics; Bacterial Toxins - metabolism; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Carp; Carps - genetics; Carps - metabolism; Carps - microbiology; Ctenopharyngodon idella; Elongation; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fish; Fish Diseases - metabolism; Fish Proteins - genetics; Fish Proteins - metabolism; Freshwater; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Profiling; Genes; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - genetics; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) - metabolism; Grasses; Life Sciences; Liver; Microbiology; Muramidase - genetics; Muramidase - metabolism; Original Research Paper; Polymerase chain reaction; Quantitative real-time PCR; Real time; Reference gene; Reference Standards; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; Validation studies
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-010-0258-0

Expression of four reference genes of grass carp, including β-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S rRNA (18S) and elongation factor-1 alpha (EF1α), was studied in tissues of normal individuals and bacteria-infected individuals. EF1α had the most stable expressions followed by 18S rRNA then GAPDH; ACTB had the least stability. After being infected with bacteria, the grass carp showed minimal changes in expression levels of EF1α in the liver and head kidney, while ACTB had the most stable expressions in spleen but the least stable in liver. EF1α is thus the optimal reference gene in quantitative real-time PCR analysis to quantitate the expression levels of target genes in tissues of grass carp.