Abstract
Human granulocyte-macrophage colony stimulating factor(hGM-CSF) is one of a family glycoprotein cytokines that have potent effects in stimulating proliferation,maturation and function of hematopoietic cells.The gene encoding hGM-CSF was PCR amplified from the plasmid pAGMT-8 and modified by addition SD sequence to promote prokaryotic expression.The resultant hGM-CSF was inserted into downstream of the strong promoter,PpsbA of expression vector pRL-439,then ligated with pDC-08 to produce the shuttle expression vector,pDC-GM.The resulting pDC-GM was transferred into the filamentous,heterocystour cyanobacterium,Anabaena PCC7120,by the tri-parental conjugation transfer method.When cells were cultivated without antibiotics for many generations,the transgenic cyanobacteria remained resistant to neomycin.PCR amplification of wild type and transgenic Anabaena cells with primers P_1 and P_2 revealed no band in wild type and plasmid free cells,whereas transgenic cells yielded a about(390 bp) band.This is the first report of expression hGM-CSF in cyanobacteria cells.
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