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Construction and Identification of hGM CSF Expressing Plasmid Which can be Stably Expressed in Mammalian Cells

Zhao Jianyang,et al (Institute of Blood transfusion,Chinese Acedemy of Medical Sciences)  
Obiective:The recombinant human glanulocyte macrophage colony stimulating factor (rhGM CSF) with natural glycosylation structure was to be obtained.Method:The eukaryocyte expressing vetor containing sRa promotor and neomycin resistance gene was used,and then it was connected to Xhol enzyme digested hGM CSF cDNA fragment,thereby pMEneo hGM CSF plasmid which could stably experss recombinant hGM CSF in mammalian cells was constructed.Results:8 of 22 pMEneo reconstructed clones contained hGM CSF cDNA fragment.Determined by enzyme digestion,the plasmids had five cis forms,one cisroasary and two trans insertions.Conclusion:The cos cells transfected by DEAE method in the mentioned above forms could traniently express the identification.The activity of hGM CSF in culture supernatant in cis form and cis rosary form was (1×10 3~1×10 4)U/ml,and there was no activity of the culture supernatant in trans insertion.
【CateGory Index】: Q343,
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