[High efficiency expression of IL-4 using modified cDNA expression vector in mammalian cells].
In order to express eukaryotic gene in eukaryotic cells, a cDNA library was constructed and screened with synthesized probe of IL-4 cDNA. The obtained cDNA fragment 739 bp in length was inserted into pi H3M plasmid and expressed (8 x 10(3) IU/ml) in COS cells. A pdB-HhIL-4 expression plasmid was constructed using pdKCR and pdBPV-1 plasmid and transfected into mouse C127 cells by calcium phosphate coprecipitation. Transfected cells exhibited altered cell morphology, could be continuously cultured in DMEM essential medium and secreted IL-4 lymphokine into 1% FCS condition culture medium. High biological activity (2 x 10(4)/ml) from the culture supernatant was obtained by FACS analysis of Fc epsilon receptor expression on B cell surface.
Wang
Z
Z
Heilongjiang Branch, CAMS, Harbin.
Tian
C
C
Liu
L
L
Xu
M
M
Lu
J
J
D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression; D015847:Interleukin-4 / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D014162:Transfection
D018076:DNA, Complementary; D004274:DNA, Recombinant; D015847:Interleukin-4