Cloning and identification of human granulocyte-macrophage colony-stimulating factor cDNA

Objective: To construct recombinant granulocyte-macrophage colony-stimulating factor(GM-CSF) retrovirus expressing vector by cloning of GM-CSF genes from human blood and construct the recombinant plasmids encoding for GM-CSF. Methods: Primers for GM-CSF were designed and synthesized according to the sequences of human GM-CSF genes derived from GenBank.The full length cDNA of GMCSF was cloned by RT-PCR techniques.The recombinant plasmids pGEM-T-GM-CSF were constructed by recombinant gene techniques.Results: The length of RT-PCR product coincided with that of authors′ anticipation(456bp),and the recombinant plasmid was confirmed by XbaⅠ /NotⅠ restriction enzyme digesting.The sequencing result of the cDNA was identical to the sequence of GM-CSF cDNA in GenBank,and the full length cDNA of human GM-CSF was successfully inserted into the vector of pGEM-T.Conclusion: The successful cloning of human GM-CSF cDNA,as well as construction of its retrovirus expressing vector enables us to further investigate the role of GM-CSF in tumor immunogene therapy.