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生物工程学报 2006
Construction and Characterization of a Recombinant Fowlpox Virus Co-expressing F, HN Genes of Newcastle Disease Virus and gB Gene of Infectious Larygnotracheitis Virus
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Abstract:
The Fusion (F) and Haemagglutinin-Neuraminidase (HN) genes of Newcastle disease virus (NDV) and the glycoprotein B (gB) gene of infectious laryngothracheitis virus (ILTV) as well as a LacZ reporter gene were all inserted into a nonessential gene of fowlpox virus (FPV) 017 strain by homologous recombination. The NDV and ILTV genes were each under the control of a fowlpox virus immediate early/late promoter (LP2EP2) while the LacZ reporter gene expression cassette was regulated by a P11 late promoter. A recombinant FPV harboring the F,HN and gB genes as well as the LacZ gene,designated as rFPV-F/HN/gB/LacZ,was obtained after ten cycles of blue plaque purification. The presence of the NDV and ILTV genes was confirmed by PCR. The expression of the recombinant proteins in rFPV-F/HN/gB/LacZ were characterized by Western blot (F and gB proteins) and indirect immunofluorescence test (F,HN and gB proteins). The results demonstrated that all four foreign proteins,which were encoded within a 10kb gene fragment,could be expressed authentically and efficiently. Compared to the parental virus,rFPV-F/HN/gB/LacZ showed no obvious difference with respect to virus replication and cytopathogenic effects in chicken embryo fibroblasts (CEF) cell culture. Overall,our work suggests that FPV can be a useful live virus vector for the expression of multi- foreign genes against multiple avian pathogens.
