Construction of recombinant fowlpox viruses as vectors for poultry vaccines.
Plasmid vectors have been constructed which allow the construction of infectious fowlpox virus (FPV) recombinants expressing foreign genes. The foreign genes were inserted within the thymidine kinase (TK) gene of FPV contained in these vectors. To facilitate the selection of recombinants the Escherichia coli xanthine guanine phosphoribosyl transferase (Ecogpt) gene was developed as a dominant selectable marker. This marker operates in a wide variety of cell types and obviates the need for TK- cell lines for selection of TK- recombinants when foreign genes have been inserted within the TK gene of FPV. The general approach adopted was to construct plasmid vectors in which the FPV TK was interrupted by the Ecogpt gene under the control of a poxvirus promoter in tandem with a gene of interest under the control of another poxvirus promoter. Selection of viruses expressing the Ecogpt gene simultaneously selects for recombinants carrying both the Ecogpt gene and the gene of interest. Using this approach a series of plasmid vectors was constructed in which the FPV TK gene was interrupted by the Ecogpt gene under the control of the P7.5 vaccinia virus promoter in tandem with the A/PR/8/34 haemagglutinin gene under the control of the PL11 vaccinia virus promoter. A recombinant FPV constructed using these plasmids had the expected genome arrangement, expressed influenza haemagglutinin, and induced haemagglutination-inhibiting antibodies when inoculated into chickens. These techniques should allow the construction of a variety of recombinant FPVs expressing poultry vaccine antigens. Such recombinants should be a very cost-effective means of delivering vaccines to poultry.
Boyle
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Commonwealth Scientific and Industrial Research Organisation, Australian Animal Health Laboratory, Geelong, Victoria.
Coupar
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BE
D000818:Animals; D000941:Antigens / Q000302:isolation & purification*; D002642:Chick Embryo; D002645:Chickens / Q000276:immunology*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000276:immunology; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005587:Fowlpox virus / Q000201:enzymology / Q000235:genetics*; D005819:Genetic Markers; D005822:Genetic Vectors; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology; D009980:Influenza A virus / Q000235:genetics; D010430:Pentosyltransferases / Q000235:genetics; D010957:Plasmids; D011212:Poxviridae / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013937:Thymidine Kinase / Q000235:genetics; D014614:Vaccines, Synthetic / Q000276:immunology / Q000302:isolation & purification*; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology
D000941:Antigens; D004274:DNA, Recombinant; D004279:DNA, Viral; D005819:Genetic Markers; D006389:Hemagglutinins, Viral; D014614:Vaccines, Synthetic; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase; D013937:Thymidine Kinase