title: Identification of suitable reference genes for quantitative real-time PCR normalization in blotched snakehead Channa maculata.
abstract: A systematic study was conducted to identify reliable reference genes for normalization of gene expression analysis in the blotched snakehead Channa maculata under normal physiological conditions. Firstly, the partial complementary (c)DNA of nine candidate reference genes (actb, tmem104, ube2l3, ef1α, churc1, tmem256, rpl13a, sep15 and g6pd) were cloned from C. maculata. The expression levels of these genes were then assessed in embryos of different developmental stages and various tissue types of adult fish using quantitative real-time (qrt-)PCR. RefFinder algorithm was used to evaluate the expression stability of these genes based on their cycle-threshold (Ct) values in the qrt-PCR analysis. Results showed that there was no single best reference gene for all stages of embryos and adult tissues tested. Furthermore, it was found that, among the nine candidate genes tested, actb and tmem104 were the most stable reference genes across adult tissue types, while sep15 and tmem256 were the most stable ones across developmental stages of embryos. These stable reference genes are recommended for normalization of gene expression analysis in C. maculata.
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.|College of Fisheries and Life Science, Shanghai Ocean University, 999 Huchenghuan Road, Shanghai, 201306, China.
authors.forename: H
authors.initials: H
authors.lastname: Mao
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.|College of Fisheries and Life Science, Shanghai Ocean University, 999 Huchenghuan Road, Shanghai, 201306, China.
authors.forename: K
authors.initials: K
authors.lastname: Chen
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.
authors.forename: X
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Luo
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.
authors.forename: J
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.
authors.forename: W
authors.initials: W
authors.lastname: Li
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.|College of Fisheries and Life Science, Shanghai Ocean University, 999 Huchenghuan Road, Shanghai, 201306, China.
authors.forename: X
authors.initials: X
authors.lastname: Wu
authors.affiliation: Pearl River Fisheries Research Institute, Chinese Academic of Fisheries Sciences, Guangzhou, 510380, China.|College of Fisheries and Life Science, Shanghai Ocean University, 999 Huchenghuan Road, Shanghai, 201306, China.
authors.forename: H
authors.initials: H
authors.lastname: Xu
mesh_terms: D000818:Animals; D004625:Embryo, Nonmammalian / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism; D005399:Fishes / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods / Q000662:veterinary; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards
chemical_list: D029941:Fish Proteins
keywords: blotched snakehead; housekeeping genes; qrt-PCR