关键词: 粒细胞巨噬细胞集落刺激因子, 基因, 克隆, 分子

Abstract: ［ABSTRACT］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 GMCSF 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.

Key words: Genes, Cloning, KEY WORDS］Granulocytemacrophage colonystimulating factor, molecular