Construction and expression of HSC70 gene eukaryotic expression plasmid

Objective To construct the HSC70 (heat shock cognate protein 70) eukaryotic expression plasmid and express it in HepG2 cells. Methods The HSC70 gene was amplified by RT-PCR from human QZE hepatocellular cells and cloned into the eukaryotic expression vector pcDNA(TM)3.1/V5-HisA,then the plasmid pcDNA-HSC70 was constructed. After identified by restriction endonuclease and sequencing,the plasmid was transfected into HepG2 cells to express HSC70-His fusion protein. The expression of HSC70-His was analyzed by immunocytochemical technique. Results 1960 bp fragment of HSC70 cDNA sequence was obtained by RT-PCR,and it was consistent with the known HSC70 sequence in GenBank. After the plasmid pcDNA-HSC70 being transfected into HepG2 cells,the fusion protein HSC70-His was instantaneously expressed in some of cells. Conclusion Recombinant eukaryotic expression plasmid pcDNA-HSC70 is successfully constructed and instantaneously expressed in HepG2 cells.