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 it s expression in COS-7 cells. Methods 　S100A13 cDNA was amplified by RT-PCR f rom papillary thyroid carcinoma tissues, Fresh PCR product s was cloned into p GEM-T vector and sequenced. Then the recombinant ORF was subcloned into pcDNA 3. 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 western2blot, 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 const ructed and expressed. The subcellular localization of S100A13 is in the whole cytoplasm in COS-7 cells .