Adaptation of a retrovirus as a eucaryotic vector transmitting the herpes simplex virus thymidine kinase gene

Journal Article · Thu Apr 01 04:00:00 EST 1982 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.2.4.426· OSTI ID:5963927

Tabin, C J; Hoffman, J W; Goff, S P; Weinberg, R A

The authors investigated the feasibility of using retroviruses as vectors for transferring DNA sequences into animal cells. The thymidine kinase (tk) gene of herpes simplex virus was chosen as a convenient model. The internal BamHI fragments of a DNA clone of Moloney leukemia virus (MLV) were replaced with a purified BamHI DNA segment containing the tk gene. Chimeric genomes were created carrying the tk insert on both orientations relative to the MLV sequence. Each was transfected into TK/sup -/ cells along with MLV helper virus, and TK/sup +/ colonies were obtained by selection in the presence of hypoxanthine, aminopterin, and thymidine (HAT). Virus collected from TK/sup +/-transformed, MLV producer cells passed the TK/sup +/ phenotype to TK/sup -/ cells. Nonproducer cells were isolated, and TK/sup +/ transducing virus was subsequently rescued from them. The chimeric virus showed single-hit kinetics in infections. Virion and cellular RNA and cellular DNA from infected cells were all shown to contain sequences which hybridized to both MLV- and tk-specific probes. The sizes of these sequences were consistent with those predicted for the chimeric virus. In all respects studied, the chimeric MLV-tk virus behaved like known replication-defective retroviruses. These experiments suggest great general applicability of retroviruses as eucaryotic vectors.

• Research Organization: Center for Cancer Research and Dept. of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139
• OSTI ID: 5963927
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 2:4; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

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550300 -- Cytology
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
AROMATICS
AZAARENES
AZINES
BIOLOGICAL ADAPTATION
BIOLOGICAL MODELS
CELL CULTURES
CELL TRANSFORMATIONS
CHIMERAS
CLONING
DISEASES
DNA SEQUENCING
DNA-CLONING
ENZYME ACTIVITY
ENZYMES
ETIOLOGY
EXPERIMENT PLANNING
GENES
GENOME MUTATIONS
HERPES SIMPLEX
HETEROCYCLIC COMPOUNDS
HYBRIDIZATION
HYDROXY COMPOUNDS
HYPOXANTHINE
INFECTIOUS DISEASES
LEUKEMIA VIRUSES
MICROORGANISMS
MOLECULAR BIOLOGY
MOSAICISM
MUTATIONS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PLANNING
PURINES
PYRIMIDINES
RIBOSIDES
RIBOSOMAL RNA
RNA
SKIN DISEASES
STRUCTURAL CHEMICAL ANALYSIS
STRUCTURAL MODELS
STRUCTURE-ACTIVITY RELATIONSHIPS
THYMIDINE
TRANSFERASES
VIRAL DISEASES
VIRULENCE
VIRUSES