Cloning, genomic structure and expression analysis of ubc9 in the course of development in the half-smooth tongue sole (Cynoglossus semilaevis)

The small ubiquitin-like modifier (SUMO) pathway is an essential biological process in eukaryote, and Ubc9 is an important E2 conjugating enzyme (UBE2) for SUMO pathway and plays a critical role in cellular differentiation, development and sex modification in various species. However, the relationship between Ubc9 and sex modification and development in fish remains elusive. To elucidate the impact of Ubc9 on sex modification and development, the full length of the cDNA and genomic sequence was cloned from half-smooth tongue sole, Cynoglossus semilaevis. Real-time quantitative RT-PCR demonstrated that ubc9 was ubiquitously expressed in different tissues, and the expression levels varied in the different stages of embryonic and gonadal development. In addition, the expression level was significantly higher in the temperature-treated females than the normal females and males. Moreover, the PET-32-Ubc9 plasmid was constructed and the recombinant protein was expressed in Escherichia coli. Follistatin gene expression was initially up-regulated and FSE genes (cyp19a1a, ctnnb1, foxl2) were initially down-regulated after the injection of Ubc9 protein, prior to 96 h eventually recovered to normal levels. Taken together, the results show that Ubc9 is involved in embryogenesis, gametogenesis and sex modification, and exerts an effect on gene expression.