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), Vol. 3:12
- Country of Publication:
- United States
- Language:
- English
Similar Records
Expression of human factor IX in rat capillary endothelial cells: Toward somatic gene therapy for hemophilia B
Isolation and characterization of NIH 3T3 cells expressing polyomavirus small T antigen
Related Subjects
GENE REPRESSORS
PHOSPHOTRANSFERASES
DNA-CLONING
GENES
BIOLOGICAL MARKERS
CHIMERAS
GENE AMPLIFICATION
GENE MUTATIONS
GENE RECOMBINATION
HERPES SIMPLEX
IN VIVO
LEUKEMIA VIRUSES
NEOMYCIN
PHENOTYPE
PLASMIDS
RATS
RECOMBINANT DNA
SIMIAN VIRUS
THYMIDINE
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
AZINES
CELL CONSTITUENTS
CLONING
DISEASES
DNA
DRUGS
ENZYMES
HETEROCYCLIC COMPOUNDS
INFECTIOUS DISEASES
MAMMALS
MICROORGANISMS
MOSAICISM
MUTATIONS
NUCLEIC ACIDS
NUCLEOPROTEINS
NUCLEOSIDES
NUCLEOTIDES
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PHOSPHORUS-GROUP TRANSFERASES
PROTEINS
PYRIMIDINES
RIBOSIDES
RODENTS
SKIN DISEASES
TRANSFERASES
VERTEBRATES
VIRAL DISEASES
VIRUSES
550400* - Genetics
550200 - Biochemistry