Abstract: To induce the expression of FMDV receptor integrinβ6 subunit ligand2binding domain in prokaryocyte and purify the recombinant protein.The fragment coding ovine integrinβ6 ligand-binding domain was amplified by PCR from the recombinant plasmid pGEM-β6.After doubly digested with EcoR Ⅰand SalⅠ,theβ6LBD fragment was subcloned into prokaryotic expression vector pPROEX^TMHTb which was doubly digested with same enzymes.The recombinant expression plasmid pP_RO/β6LBD was constructed and transformed into E.coli BL21(DE3) and induced by IPTG.The expression of fusion protein was detected by SDS-PAGE and purified by Ni-NTA His.Bind resins.The expressed product was identified by ELISA and Western blot assay.SDS-PAGE demonstrated that the fusion protein pP_RO/β6LBD was expressed efficiently as inclusion body in E.coli with the expected molecular weight of 33 kDa.ELISA and Western blot assays showed that the recombinant protein has the good antigenicity and specificity,which will lay a foundation for further research on distribution and the role of the ovine integrinβ6 subunit in FMDV infection.

Key words: FMDV, Receptor, Integrinβ6 subunit, Ligand2binding domain, Prokaryotic expression

CLC Number: 

• Q785