Transfection and expression of murine interleukin-12 gene in B16F10 melanoma cells

AIM:To study the expression of murine interleukin-12(mIL-12) gene in B16F10 melanoma cells.METHODS:mIL-12 gene was inserted into pcDNA3.1 eukaryotic expression vector by DNA recombination and then transfected into B16F10 melanoma cells by electroporation. The integration and expression of mIL-12 gene in B16F10 melanoma cells were detected by PCR, RT-PCR and Western blot after positive cell clones had been screened.RESULTS:mIL-12 gene was confirmed to have been transfected and expressed in B16F10 cells on three levels of DNA, mRNA and protein.CONCLUSION:mIL-12 gene can successfully be transfected and expressed in B16F10 melanoma cells in vitro, which lays a foundation for the further investigation of mIL-12 gene tumor vaccine.