Molecular cloning and expression of chemokine IP-10 and detection of its immunocompetence

In order to study the biological characteristics of swine chemokine and the effect of Interferon-inducible Protein (IP-10) on the immunocompetence of vaccine, swine IP-10 gene was amplified from Spleen by RT-PCR, and the fragment was cloned into the eukaryotic expression plasmid to construct recombinant plasmid pcDNA3.1-IP-10. Then the plasmid pcDNA3.1-IP-10 was transfected into HEK293T cell. The fusion 6×HIS-IP-10 protein was expressed. The chemotaxis of IP-10 protein was measured by transwell migration assay on spleen lymphocyte. And the animals were immunized with the vaccine which the active protein was as adjuvant. Then the regulation effect of IP-10 protein on immunization was determined. The construction of IP-10 recombinant vector was proved by enzyme digestion and sequencing. The IFA and Western blotting shows that HEK293T cell could produce IP-10 protein successfully. The recombinant of IP-10 protein has chemotaxis on spleen lymphocyte, and it can have function of immunologic enhancement. The IP-10 recombinant protein can be the promising immune adjuvant to be used in practice for its biological function of chemotaxis.