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| Construction and identification of recombinant adenovirus coexpression PRRSV GP5 gene and porcine interferon-γ gene |
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Abstract In this study, the GP5 gene of PRRSV and the porcine IFN-γ were amplified by PCR, and ligated by IRES sequence ,then cloned into the transfer vector pAdenoVator-CMV5-IRES-GFP.After co-transformation of PmeI-linearized recombinant plasmid plRES-GP5-IFN-γ and the bone vector pAdEasy-1 into Escherichia coli bacteria strain BJ5183, recombinant plasmid containing GP5 gene and IFN-γ(pAd-GP5- IFN-γ) was obtained and identified with PCR.Upon transfection of PacI-linearized plasmid pAd-GP5- IFN-γ in 293 cell line, a recombinant adenovirus was obtained and named as rAd- GP5- IFN-γ with viral titer of 2×109 TCID50/ml. The expression of the E protein and IFN-γ in the 293 cells infected with rAd- GP5- IFN-γ was confirmed with specific antibodies to E protein and IFN-γ byWestern blotting. It indicated that a stable and safety replication defective recombinant adenovirus rAd-GP5-IFN-γ was produced successfully The recombinant adenovirus might be an attractive candidate vaccine for preventing the disease of PRRS.
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Received: 09 June 2006
Published: 10 May 2010
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