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[Gene cloning and expression of human soluble interleukin-6 receptor in insect cells]
1Department of Biotechnology, Institute of Medicinal Biotechnology, CAMS, PUMC, Beijing 100050, China.
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.