Molecular Cloning, Sequence Structure and Expression Analysisof Gonadal Aromatase Gene in Puffer Takifugu obscures
GAO Yingying1,2, LIU Xinfu1,3, HU Peng4, HUANG Bin1,3, LIU Bin1,3
1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; 2.College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306, China; 3.Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China; 4.Neijiang Normal University, Neijiang 641100, China
Abstract：The gonadal aromatase gene (cyp19a) was a kind of gene that regulates the sex hormone balance. The purpose of this study was to preliminarily investigate the role of gonadal aromatase gene in gonad development and sex differentiation in puffer Takifugu obscurus, and we successfully cloned the full-length cDNA sequence of cyp19a in puffer through RACE and Real-Time PCR technology. The full length of cyp19a sequence was 1786 bp, encoding 514 amino acids, and sharing the maximal homology with the genes of puffer T. rubripes and T. niphobles. The amino acid sequence analysis showed that the protein encoded by cyp19a was hydrophilic and stable protein, but there is no signal peptide sequence. There were 38 phosphorylation sites, 4 N-glycosylation sites, 2 transmembrane domains in the sequence. Meanwhile, the multiple amino acid sequence alignment showed that the amino acid sequence of gonadal aromatase was very conservative in both mammalian and fish, and contained some functional conservative regions such as the transmembrane region, I-helix region, Ozol′s peptide region, aromatase-specific conserved region and heme binding region. Furthermore, expression analysis showed that the cyp19a was highly abundant in ovary and muscle, but less abundant in other tissues. The expression of cyp19a in the ovary of juvenile fish at different developmental stages showed a trend of increasing first and then decreasing rapidly, without in the testis. The findings indicated that cyp19a may be involved in the development of ovarian after gonad differentiation, maintenance of female characteristics and development of other tissues in female puffer.
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2020, Vol. 39 
