Construction of Recombinant Plasmid SV40T/pcDNA3.1 and Its Expression in Hepatocytes

To construct a recombinant plasmid SV40T/pcDNA3.1 for the construction of immortalization hepatocytes line, pcD2 plasmid was digested by restriction enzyme BamH Ⅰ to harvest the SV40T DNA fragment. DNA fragment was inserted to the vector pcDNA3.1 by identification of restriction enzyme digestion. Hepatocytes were isolated and cultured according to the standard methods. Indirect immunofluorescence test was carried out using SV40T monoclonal antibody to identify its expression in hepatocytes.