Objective:To construct recombinant plasmids containing 3 T cell epitopes from ESAT-6 antigen of Mycobacterium tuberculosis and extra-cellular fragment of Flt3 ligand(FL) genes,and to express them in rat glomerular mesangial cells(GMC). Methods:The amino acid sequence of ESAT-6 antigen was analysed using predictive algorithms and 3 T cell epitopes were predicted. The oligonucleotide encoding and the linker Ala-Ala-Tyr(AAY) were synthesized and inserted into the bicistronic vector pIRES and pIRES-FL. The recombinant plasmids were transfected into GMC cells,the recombinant proteins expressing in GMC cells was examined by Western blot. Results:The recombinant plasmids were verified by sequencing and the recombinant fusion proteins were identified by Western blot. Conclusion:The multiepitope DNA vaccine from ESAT-6 antigen of Mycobacterium tuberculosis was constructed and expressed successfully.