Validation of reference genes of grass carp Ctenopharyngodon idellus for the normalization of quantitative real-time PCR.
Expression of four reference genes of grass carp, including beta-actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), 18S rRNA (18S) and elongation factor-1 alpha (EF1alpha), was studied in tissues of normal individuals and bacteria-infected individuals. EF1alpha 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 EF1alpha in the liver and head kidney, while ACTB had the most stable expressions in spleen but the least stable in liver. EF1alpha 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.
Ye
Xing
X
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of Tropical & Subtropical Fish Breeding & Cultivation, CAFS, Guangzhou 510380, People's Republic of China. gzyexing@163.com
Zhang
Lili
L
Dong
Haiyan
H
Tian
Yuanyuan
Y
Lao
Haihua
H
Bai
Junjie
J
Yu
Lingyun
L
D000818:Animals; D001424:Bacterial Infections / Q000378:metabolism / Q000662:veterinary*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism; D002347:Carps / Q000235:genetics* / Q000378:metabolism* / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000378:metabolism*; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism*; D020869:Gene Expression Profiling; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics / Q000378:metabolism; D009113:Muramidase / Q000235:genetics / Q000378:metabolism; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D001427:Bacterial Toxins; C404907:Efa1 protein, E coli; D029968:Escherichia coli Proteins; D029941:Fish Proteins; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D009113:Muramidase