Construction of recombinant retroviral vector carrying herpes simplex virus thymidine kinase-1 gene and identification of its expression in HeLa cells

Objective:To construct the recombinant retrovirus carrying herpes simplex virus thymidine kinase-1(HSV-1TK) gene and to observe the expression of HSV-1TK in cervical carcinoma cell line HeLa. Methods:The HSV-1TK cDNA was digested by restriction endonuclease from plasmid pcDNA3.1/HA-myc-His(-) Z-TK and then directionally inserted into retroviral vector pLXSN to construct recombinant plasmid pLXSN-TK. The pLXSN-TK was transferred into retroviral producer cell line PA317 for packaging retrovirus and then the resultant retrovirus was applied to infected the HeLa cells. The integration of HSV-1TK gene into the HeLa cells genome was assayed by PCR. The expression of HSV-1TK gene was detected by RT-PCR and Western blot. Results:The HSV-1TK gene was integrated into the genome of HeLa cells shown by results of PCR after retroviral infection. The expression of HSV-1TK gene was confirmed by RT-PCR and Western blotting. Conclusion:The recombinant retrovirus carrying HSV-1TK gene was constructed successfully and may be a potentially effective strategy for the treatment of cervical cancer.