Aim: To construct a DNA vaccine based on surface Ig V(H) gene of tumor cells in the human B-cell lymphoma biopsy tissue.
Methods: The V(H) gene fragment was amplified by RT-PCR using Ig superfamily primers. Also, the murine monocyte chemotactic protein 3(MCP-3) cDNA was cloned. The fusion gene fragment of MCP-3 gene with V(H) gene was constructed by recombinant PCR and then cloned into the eukaryotic expression vector pcDNA3.1 to construct the DNA vaccine plasmid pcDNA3.1/MCP-V(H). The vaccine plasmid was transiently expressed in the eukaryotic cell line COS-7.
Results: The DNA vaccine plasmid was successfully constructed and expressed in COS-7 cells in the form of fusion protein MCP-V(H).
Conclusion: The DNA vaccine plasmid pcDNA3.1/MCP-V(H) is constructed and expressed successfully, which plays the foundation for further experimental research in animal model.