Transient marker stabilisation: a general procedure to construct marker-free recombinant vaccinia virus.
Recombinant vaccinia viruses based on the highly attenuated Modified Vaccinia Ankara (MVA) strain expressing HIV-1 antigen genes were constructed by a novel procedure involving the transient use of two marker genes. The selectable markers used, the Escherichia coli guanine phosphoribosyltransferase (gpt) and the beta-galactosidase (lacZ) genes, are not retained within the final recombinant virus. The transient marker stabilisation (TMS) procedure allows the generation of marker-free recombinant viruses in a series of simple plaque purification steps. HIV-1 gag pol genes were inserted into two loci of vaccinia MVA demonstrating the efficiency of the procedure.
Scheiflinger
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Immuno AG, Orth/Donau, Austria.
Dorner
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Falkner
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FG
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology*; D015685:Fusion Proteins, gag-pol / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D015488:HIV Antigens / Q000235:genetics*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured; D014616:Vaccinia virus; D001616:beta-Galactosidase / Q000235:genetics*
D015685:Fusion Proteins, gag-pol; D015488:HIV Antigens; D007041:Hypoxanthine Phosphoribosyltransferase; D001616:beta-Galactosidase