%0 Journal Article
%A Gan Ren-Bao
%A Jin Rui
%A MA Wei-Min
%A SHI Ding-Ji
%A WANG Quan-Xi
%A WEI Lan-Zhen
%T Cloning of human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene in <em>Anabaena</em> sp. PCC7120
%D 2005
%R 10.7525/j.issn.1673-5102.2005.04.013
%J Bulletin of Botanical Research
%P 436-440
%V 25
%N 4
%X 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, <em>Anabaena</em> 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 <em>Anabaena</em> cells with primers P<sub>1</sub> and P<sub>2</sub> 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.
%U https://bbr.nefu.edu.cn/EN/10.7525/j.issn.1673-5102.2005.04.013