JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY)

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Cloning and identification of human granulocytemacrophage

LI Minxiong1, CHEN Shengqiang2, LIU Qicai3, ZHANG Jianguo1   

  1. (1. Department of Otorhinolaryngology; 2. Institute of Neurosciences; 3. Experimental Medical Research Center, Second Affiliated Hospital of Guangzhou Medical College, Guangzhou 510260, Guangdong, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-04-25 Published:2006-04-25

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

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