Molecular Cloning and Expression Analyses of Foxo3a and Foxo3b Genes in Gobiocypris rarus
QIAN Jiaming1, MIN Qianwen2, DONG Ranran1, ZENG Sheng2, ZHANG Xianbo2
1. College of Animal Science, Guizhou University, Guiyang 550025, China; 2. Guizhou Fisheries Research Institute,Guizhou Academy of Agriculture Sciences, Guiyang 550025, China
Abstract:The full-length cDNA of forkhead protein O3a (Foxo3a) and forkhead protein O3b (Foxo3b) genes were cloned from the ovary of Gobiocypris rarus by using RACE. RT-PCR, qRT-PCR and in situ hybridization were employed to detect the expression of the pair of paralogous genes. A 2562 bp full-length cDNA of Foxo3a gene was abtained, containing a 1953 bp open reading frame (ORF) that encoded a protein of 650 amino acids, 242 bp 5' untranslated terminal region (UTR), 367 bp 3' UTR; a 2804 bp full-length cDNA of Foxo3b gene, containing a 1959 bp ORF that encoded a protein of 652 amino acids, 470 bp 5' UTR, 375 bp 3' UTR. Foxo3a gene was only expressed in brains and ovaries with a high level among all detected tissues. Foxo3b gene was highly expressed in brains and ovaries, and also found to be expressed in all other detected tissues. Foxo3a gene mRNA was detected in the ovaries with high levels from 2 month onward and slightly increased from 4 to 5 months. There was no significance in Foxo3b gene expression level in ovaries at all developing stages. In situ hybridization results showed that both Foxo3a and Foxo3b genes were expressed in the germ cell and follicular cell of Ⅱ, Ⅲ, and Ⅳ stage oocytes. Foxo3a gene expression was affected, but Foxo3b gene not, after the treatment by rapamycin. These findings indicate that Foxo3a and Foxo3b genes may play some roles in the oogenesis of G. rarus.
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