Retrovirus transduction: Generation of infectious retroviruses expressing dominant and selectable genes is associated with in vivo recombination and deletion events
The authors describe the generation of infectious retroviruses containing foreign genes by an in vivo recombination-deletion mechanism. Cotransfection into mouse cells of chimeric plasmids carrying a murine retrovirus 5' long terminal repeat and either the thymidine kinase (tk) gene of herpes virus or the dominant selectable bacterial gene for neomycin resistance (neo), along with a clone of Moloney murine leukemia virus, results in the generation of infectious thymidine kinase or neomycine-resistant viruses. Expression of the selectable marker in these viruses can be regulated by the homologous transcriptional promoter of the gene, by the promoter contained within the Friend spleen focus-forming virus long terminal repeat, or by the simian virus 40 early region promoter. In all cases, the rescued viruses appeared to arise by recombination in vivo with Moloney murine leukemia virus sequences, resulting in the acquisition of the Moloney 3' long terminal repeat and variable amounts of the 3' adjacent Moloney genome. In two of the thymidine kinase constructs where tk was inserted 200 base pairs downstream from the long terminal repeat, the rescued viruses acquired a large part of the murine leukemia virus genome, including the region involved in packaging genomic RNA into virions. The generation of infectious neomycin-resistant virus is associated with deletions of simian virus 40 splicing and polyadenylation sequences. These results demonstrate that nonhomologous recombination and deletion events can take place in animal cells, resulting in the acquisition or removal of cis-acting sequences required for, or inhibitory to, retrovirus infectivity.
- Research Organization:
- Ontario Cancer Institute and Dept. of Medical Biophysics, Univ. of Toronto, Toronto, Ontario M4X 1K9
- OSTI ID:
- 5877848
- Journal Information:
- Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 3:12; ISSN MCEBD
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
AZINES
BIOLOGICAL MARKERS
CELL CONSTITUENTS
CHIMERAS
CLONING
DISEASES
DNA
DNA-CLONING
DRUGS
ENZYMES
GENE AMPLIFICATION
GENE MUTATIONS
GENE RECOMBINATION
GENE REPRESSORS
GENES
HERPES SIMPLEX
HETEROCYCLIC COMPOUNDS
IN VIVO
INFECTIOUS DISEASES
LEUKEMIA VIRUSES
MAMMALS
MICROORGANISMS
MOSAICISM
MUTATIONS
NEOMYCIN
NUCLEIC ACIDS
NUCLEOPROTEINS
NUCLEOSIDES
NUCLEOTIDES
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PHENOTYPE
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PLASMIDS
PROTEINS
PYRIMIDINES
RATS
RECOMBINANT DNA
RIBOSIDES
RODENTS
SIMIAN VIRUS
SKIN DISEASES
THYMIDINE
TRANSFERASES
VERTEBRATES
VIRAL DISEASES
VIRUSES