Construction of retroviral vector containing HSV1-TK gene and its expression

Objective: To construct a retroviral vector containing TK gene and examine its expression in the lung cancer cells transfected with this gene for facilitating the study of gene therapy for lung cancer. Methods A approximately 2.4 kb TK gene segment was excised from plasmid PHSV106,and inserted into the polyclonal site of retroviral vector PLXSN. A positive linking plasmid was selected by restriction endonuclease analysis and used to transduce pulmonary cancer cell line A549 by electroporation. in which the recombination and expression of TK gene were examined by PCR and in situ hybridization. Results A positive connective plasmid was selected and verified by restriction endonuclease analysis, and the transgene-bearing cells were screened by G418, in which TK gene expression was observed. Conclusion A retroviral vector containing TK gene was sucessfully constructed, and the cells transfected with the vector are capable of expressing TK gene.