[Impact of AFP promoter on the expressions of GFP in tumor cells].
OBJECTIVE
To study the impact of AFP 5'flanking promoter (enhancer) on the expression of GFP in hepatocarcinoma cell.


METHODS
Green Fluorescent Protein (GFP) reporter gene expression plasmid pcDNA3-GFP-AFP-w under the direction of AFP 5' flanking promoter (enhancer) was constructed by recombinant DNA technology and confirmed by restriction analyses. pcDNA3-GFP-AFP-w, pcDNA3-GFP and pcDNA3 were transfected into Hela and Bel7402 cells by lipofectin and selected by G418 respectively, after amplification of the positive cell clones, expression of GFP was detected by Western blotting and quantitatively analysed by GEL Doc 2000 digital image systems.


RESULTS
The expression of GFP was lower in Bel-GFP-AFP-w than in Bel-GFP but was significantly higher than in Hela-GFP-AFP-w.


CONCLUSIONS
GFP reporter gene plasmid pcDNA3-GFP-AFP-w under the direction of the 3.1 kb AFP 5'flanking promoter (enhancer) can be expressed in HCC Bel7402 cell definitely and specifically.
Mao
C Q
CQ
Beijing Hospital, Beijing 100730, China. maocq@nic.bmi.ac.cn
Huang
C Z
CZ
Yang
S D
SD
Zhao
M
M
Fan
Y X
YX
Xu
F
F
Du
F
F
D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D008164:Luminescent Proteins / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000235:genetics*
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000509:alpha-Fetoproteins; D049452:Green Fluorescent Proteins