[Gene cloning and expression of human soluble interleukin-6 receptor in insect cells].
OBJECTIVE
To clone the human soluble interleukin-6 receptor(hsIL-6R) gene and expression in insect cell line.


METHODS
The hsIL-6R gene was cloned into plasmid pAcGP67B. After co-transfection of recombinant plasmid and wild type AcNPV DNA, the rAcNPV was confirmed by the end point dilution assay and dot blot. Then it was purified by plaque assay.


RESULTS
SDS-PAGE showed molecular weight of the expressed product was about 47,000. The expressed recombinant protein was confirmed to be specific and capable of binding its ligand IL-6 by Western blot and ligand-receptor binding assays.


CONCLUSIONS
Secretory expression of hsIL-6R gene in baculovirus expression system was indicated. The expressed product had immunological and biological activities.
Chang
B S
BS
Department of Biotechnology, Institute of Medicinal Biotechnology, CAMS, PUMC, Beijing 100050, China.
Li
Y
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006801:Humans; D019947:Receptors, Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D018076:DNA, Complementary; D019947:Receptors, Interleukin-6