Cloning, Tissue Distribution and In vitro Interaction of scp3 and dmc1 Genes in Silurus meridionalis
LIU Zhi-Hua1, DONG Ran-Ran1,*, QIAN Jia-Ming1, ZHANG Xian-Bo2
1 College of Animal Science/Institute of Special Aquatic Products, Guizhou University, Guiyang 550025, China; 2 Fishery Research Institute, Guizhou Academy of Agricultural Sciences, Guiyang 550025, China
Abstract：Synaptonemal complex 3 (SCP3) and DNA meiotic recombinase 1 (DMC1) are key factors in meiosis, which have not been reported in Silurus meridionalis. In present study, PCR, fluorescence immunohistochemistry and Western blot were used to clone and analyze the sub-cellular distribution and tissue expression of scp3 and dmc1; GST pull-down was used to verify in vitro protein-protein interaction. The results showed that the CDS of scp3 (GenBank No. MW075231) and dmc1 (GenBank No. MF770581) were 723 and 1 029 bp, encoding 240 and 342 amino acids, respectively. Homology analysis showed that the homology of S. meridionalis SCP3 and DMC1 to Tachysurus fulvidraco were as high as 89.17% and 97%, respectively. In phylogenetic analysis, S. meridionalis SCP3 and DMC1 were clustered with fish, and then clustered with tetrapod. Reverse transcription-PCR (RT-PCR) results showed that scp3 and dmc1 were only specifically expressed in ovary. The results of qRT-PCR showed that relative expression of scp3 and dmc1 had the highest level in 55 dah (days after hatching), which were significantly higher than that of 30 and 90 dah. The results of fluorescence immunohistochemistry showed strong positive signals of SCP3 in nucleus and cytoplasm of oocytes, and strong positive signal of DMC1 in the nucleus of oocytes. GST pull-down assay showed that SCP3 interacted with DMC1 in vitro. This study could provide basic data for further study on the function of SCP3 and DMC1 during meiosis, and reference for artificial breeding of S. meridionalis to obtain high-quality eggs.
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