[Cloning, expression and identification of IL-1ra-Fcepsilon fusion gene].
Both interleukin-1 and IgE are important in the pathogenic mechanism of the allergy asthma. cDNA of interleukin-1receptor antagonist (IL-1ra) and IgE were cloned and a prokaryotic expression vector IL-1ra-Fcepsilon/pBV220 was constructed. The vector was transformed into Escherichia coli BL21(DE3). The fusion protein was expressed successfully in the form of inclusion body. The recombination protein of IL-1ra-Fcepsilon was highly purified by chromatography of gel filtration and ion exchange, which was identifited by Western blotting. The cell assay showed that the activity of IL-1ra-Fcepsilon was as high as IL-1ra in vitro after refolding. The pharmacokenetic profile of IL-1ra-Fcepsilon and L-1ra was analyzed, and the half time of IL-1ra-Fcepsilon is 4.78 times than that of IL-1ra.
Liu
Zhongcheng
Z
College of Pharmacy, Hebei University, Baoding 071002, China.
Zou
Minji
M
Wang
Yuanyuan
Y
Wang
Jiaxi
J
Xu
Donggang
D
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D050939:Gene Fusion; D007073:Immunoglobulin E / Q000235:genetics* / Q000378:metabolism; D007141:Immunoglobulin Fc Fragments / Q000235:genetics* / Q000378:metabolism; D053590:Interleukin 1 Receptor Antagonist Protein / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism
D007141:Immunoglobulin Fc Fragments; D053590:Interleukin 1 Receptor Antagonist Protein; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E