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Custom Vector Construction> | Transient transfection in COS-7 cells was conducted as previously described in (Aizen et al., 2016). Co-transfection of 10 ng of the UAS reporter vector pGL4.35[luc2P/9XGAL4UAS/Hygro] (Promega) and 20 ng of the GAL4-fusion plasmids pFN26A (BIND) hRluc-neo Flexi® Vector (Promega) containing the GAL4 binding domain fused to: iDmrt1 open reading frame (iDmrt1); iDmrt1C′ corresponding to amino acids (aa) 157–504 (iDmrt C′); iDMY open reading frame (iDMY); iDMY C′, corresponding to aa 143-179 (iDMY C′); and empty vector (plasmid constructs generated by Genscript) | Get A Quote |
Sex determination pathways are extensively diverse across species, with the master sex-determinants being the most variable element. Despite this, there is a family of DM-domain transcription factors (Dmrts), which hold a highly conserved function in sexual development. This work is the first to describe a heterogametic sex-linked Dmrt in an invertebrate species, the Eastern spiny lobster, Sagmariasus verreauxi. We have termed the Ylinked, truncated paralogue of the autosomal iDmrt1, Sv-iDMY. Considering the master sex-determining function of both DMY in medaka and DM-W in frog, we hypothesised a similar function of Sv-iDMY. By conducting temporal expression analyses during embryogenesis we have identified a pu... More