Expression and Localization of pEGFP-C3/Tsarg1 Fusion Protein in GC-1 spg Cells

Objective To construct eukaryotic expression vector pEGFP-C3/Tsarg1 and to observe the subcellular localization in GC-1 spg cells.Methods The open reading frame(ORF)of Tsarg1 was amplified from rat testis by RT-PCR.The PCR products were cloned into pUCm-T vectors and sequenced.Then the cDNA fragment was subcloned into pEGFP-C3 vector and the positive clones were selected.The recombinant plasmid pEGFP-C3/Tsarg1 was transfected into the GC-1 spg cells using liposome.The expression and localization of the fusion protein were observed under fluorescent microscope.Results The eukaryotic expression plasmid of pEGFP-C3/Tsarg1 was successfully constructed.EGFP-Tsarg1 fusion protein was expressed in the GC-1 spg cells and mainly presented in the cytoplasm after transfection with pEGFP-C3/Tsarg1.Conclusions The recombinant eukaryotic expression vector of pEGFP-C3/Tsarg1 has been constructed successfully and GFP tagged Tsarg1 is immediately expressed in the GC-1 spg cells and localized in the cytoplasm,which lay the foundation for further studying the biological functions of Tsarg1.