Abstract: Objective: To amplify the CD and TK gene fragment and analyze the DNA sequence, and then to construct eukaryotic expression plasmids pIRES-CD and pIRES-TK and determine their expression in ACC-2 cells. Methods: The primers of CD and TK were designed and composed at the base of the CD and TK's DNA sequence. CD and TK genes were cloned by PCR and then the eukaryotic expression plasmids pIRES-CD and pIRESTK were constructed and transfected into ACC-2 cells by electroporation. The transfection and transcription of CD and TK were successfully demonstrated by RT-PCR. Results: The cloned CD and TK gene segments accorded with those reported in the GeneBank. The recombination expression vector contained CD and TK genes by enzyme restriction. RT-PCR analysis demonstrated that CD and TK genes effectively expressed in ACC-2 cells. Conclusion: The CD and TK genes are successfully amplified and the sequence is analyzed. The pIRES-CD and pIRES-TK expression plasmids are successfully constructed and expressed in ACC-2 cells, which offers a pathway for research on gene therapy of tumors.

Key words: Cytosine deaminase, Thymidine kinase, Internal ribosome entry site, Endonuclease, Adenoid cystic carcinoma