Amplification of mouse GM-CSF gene and its expression in HEK293T cells

Granulocyte and macrophage colony-stimulating factor(GM-CSF) is an important cytokine in animal immune system and is also a biological adjuvant.To express GM-CSF in a secretory manner in eukaryotic cells,the mouse GM-CSF cDNA was amplified with RT-PCR from mouse spleen and was inserted into pcDNA3.1A expression vector to construct its eukaryotic expression vector,pcDNA3.1-GMCSF.The transient expression of GM-CSF was performed in HEK293T cells transfected via liposome mediation.The results showed that the amplified GM-CSF gene sequence was consistent with that published in GenBank.The recombinant mouse GM-CSF was detected by Western-blot in the culture medium of the transfected HEK293T cells,which indicates that the GM-CSF gene could be expressed and secreted in the cells.Those results provide the necessary conditions for further study on the GM-CSF adjuvant functions in animal vaccine,especially in animal DNA vaccine.