Construction of Eukaryotic Express Vector with Human S100A13 Gene and Its Expression in COS-7 Cells

Objective To obtain eukaryotic express vector of human S100A13 gene and detect its expression in COS-7 cells. Methods S100A13 cDNA was amplified by RT-PCR from papillary thyroid carcinoma tissues,Fresh PCR products was cloned into pGEM-T vector and sequenced. Then the recombinant ORF was subcloned into pcDNA3.1/NT-GFP-topo vector and sequenced. The recombinant vector was transfected into COS-7 cells by lipofectamine,the expression level of S100A13 mRNA and protein were identified by RT-PCR and western-blot,respectively.Results S100A13 gene was identified to be iserted into pcDNA3.1/NT-GFP-topovector correctly and the recombinant vector expressed S100A13 mRNA and protein stably.The green fluorescence protein was expressed in the whole cytoplasm in COS-7 cells. Conclusion The eukaryotic express vector containing human S100A13 gene was successfully constructed and expressed. The subcellular localization of S100A13 is in the whole cytoplasm in COS-7 cells .