title: Multiple-cloning-site plasmids for the rapid construction of recombinant poxviruses.
abstract: Plasmid vectors containing multiple cloning sites suitable for the rapid insertion of protein-coding sequences into poxviruses have been constructed. They are based on pUC plasmids and carry the thymidine kinase (TK) gene of vaccinia virus interrupted by a vaccinia virus promoter. Six unique restriction enzyme sites (BamHI, SalI/HincII, PstI, HindIII, EcoRI), located within 40 bp of vaccinia virus promoters transposed from the HindIII-F or HindIII-C fragment of the vaccinia virus genome, allow rapid insertion of foreign-protein-coding sequences into these plasmids. Such plasmids can be used to construct recombinant poxviruses expressing foreign proteins using marker-rescue recombination techniques and selection for TK negative viruses. Vaccinia viruses expressing the haemagglutinin (HA) gene of swine influenza virus, A/NJ/11/76 (H1N1), have been constructed.
mesh_terms: D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D005822:Genetic Vectors; D006389:Hemagglutinins, Viral / Q000235:genetics; D009980:Influenza A virus / Q000235:genetics; D009876:Operon; D010957:Plasmids; D011212:Poxviridae / Q000235:genetics*; D011995:Recombination, Genetic; D013937:Thymidine Kinase / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics
chemical_list: D004279:DNA, Viral; D006389:Hemagglutinins, Viral; D013937:Thymidine Kinase