Package and expression of lentivirus encoding hTERT gene

Objective To construct a lentiviral vector encoding human telomerase reverse transcriptase(hTERT)genes.Methods A fragment of the hTERT gene was amplified by RT-PCR and was inserted into the plasmid L166.The three plasmids L166-hTERT,L205 and L311 co-transfected the virus packaging cell line 293T using the CaCl2 method. Then the cell supernatant was harvested and virus titration was determined. Also the expression of hTERT in 293T cells infected by the lentivirus was determined by using the immunocytochemistry method.Results DNA sequencing demonstrated that the recombinant plasmid L166-hTERT was successfully constructed.The titration of the un-concentrated lentivirus was 5.25×106?IU/mL.hTERT polypeptid immunocytochemistry staining proved positive for the hTERT gene in infected 293T cells.Conclusion Lentivirus encoding hTERT was successfully packaged and hTERT could express in 293T cells for more than 2 months.