[Construction and identification of recombinant fowlpox virus for expressing haemagglutinin-neuraminidase glycoprotein of Newcastle disease virus strain F48E8].
For the construction of transfer vector pFGHN1175-1, the gene encoding haemagglutinin-neuraminidase(HN) glycoprotein of newcastle disease virus(NDV) strain F48E8 was removed from plasmid pGEMHN, and inserted into the HindIII site of insertion vector pFG1175-1, downstream of P7.5 promotor. Chicken embryo fibroblast (CEF) cell cultures which had been infected with fowlpox virus (FPV) Chinese vaccine strain 282E4 for 3-4 hours were transfected with pFGHN1175-1 plasmid DNA by liposomal transfection. Recombinant FPV with blue plaques were selected and purified 3 times in CEF cell cultures overlaid agarose containing X-gal. PCR analysis and DNA dot-blotting hybridization assay indicated that the HN gene had inserted into the FPV genomic DNA. The expression of the NDV HN gene in recombinant FPV was confirmed by indirect immunofluorescence assay with specific monoclonal antibody.
Liu
W
W
Department of Veterinary Medicine, Yangzhou University, Yangzhou 225009.
Wu
Y
Y
Jiang
Y
Y
Zhang
R
R
Liu
X
X
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D005347:Fibroblasts / Q000821:virology; D005587:Fowlpox virus / Q000235:genetics*; D005822:Genetic Vectors; D015707:HN Protein / Q000096:biosynthesis / Q000235:genetics*; D009522:Newcastle disease virus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014162:Transfection
D015707:HN Protein