Expression and functional analysis of Kcmf1 gene in the testis of Bombyx mori and Spodoptera litura

The Kcmf1 gene belongs to the E3 ubiquitin ligase family and is related to the organ development and oncogenesis in mammals. However, the function of Kcmf1 in the reproductive systems of mammals and insects remains unclear. In this paper, we obtained the ORFs of Bombyx mori BmKcmf1 (1110 bp) and Spodoptera litura SlKcmf1 (966 bp) through PCR method. Bioinformatics analysis showed that both BmKCMF1 and SlKCMF1 proteins contained conserved zinc-binding domains and C₂H₂ type zinc finger domains. Moreover, we found that BmKcmf1 and SlKcmf1 were expressed in the testis of B. mori and S. litura at different developmental stages by real-time quantitative PCR (qRT-PCR) and their expression varied with developmental stages, suggesting that BmKcmf1 and SlKcmf1 could be related to testis development. The BmKCMF1 and SlKCMF1 were overexpressed in the BmN cells and Sl221 cells, respectively, the mRNA levels of genes related to spermatogenesis such as Vasa, Hop and Dpp changed significantly. In conclusion, our results suggested that BmKcmf1 and SlKcmf1 genes might participate in the process of testis development and spermatogenesis of B. mori and S. litura.