关键词: hGM-CSF基因, 三亲接合转移, 转基因蓝藻, 鱼腥藻7120

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 P₁ and P₂ 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.

Key words: Human granulocyte-macrophage colony stimulating factor, triparental conjugative transfer, transgenic cyanobacterium, Anabaena sp.PCC7120