Construction Eukaryotic Express Vector Of Human S100A13 Gene And Reasch Its Biological Characteristics

Objective To obtain eukaryotic express vector of human S100A13 gene and detect its expression in COS-7 and HepG2 cells. Then to reasch its biological characteristics and sensitivity to anticancer drugs .Methods S100A13 cDNA was amplified by RT-PCR from papillary thyroidcarcinoma tissues,Fresh PCR products was cloned into PGEM-T vector and sequenced. Then the recombinant ORF tagged with six histidine at 5'terminal was subcloned into PcDNA3.1/NT-GFP-topo vector and sequenced. The recombinant PcDNA3.1/NT-GFP-S100A13 was transfected into COS-7 and HepG2 cells by lipofectamine, the mRNA expression level of S100A13 and FGF1 were indentified by RT-PCR,respectively. And the expression level of S100A13 protein were indentified by Western-blot.The cell proliferation status were examined by MTT technical.Then cells were treated with 5-Fu.The cell apoptosis rates were detected by flow cytometry.Results S100A13 gene tagged with six histidine at 5' terminal was indentified to be inserted into PcDNA3.1/NT-GFP- vector correctly.and the recombinant vector expressed S100A13 mRNA and protein stably.S100A13 is expressed in the cell plasm of COS-7 cells while in the whole cell of HepG2 cells, the express level of FGF1 is higher in cells transfected with recombinanted S100A13 than blank control groups and positive control groups in S100A13 stably expressed cell lines.But there were no significant difference between recombinant groups and control groups in cell growth trend and its sensitivity to anticancer drugs .Conclusion The eukaryotic express vector containing human S100A13 gene wassuccessfully constructed and expressed in COS-7 and HepG2 cells. The subcellular localization of S100A13 was different in COS-7 and HepG2 cells . S100A13 could...