Deoxyribonucleic acid-mediated gene transfer in mammalian cells: Molecular analysis of unstable transformants and their progression to stability
To elucidate mechanisms involved in deoxyribonucleic acid-mediated gene transfer, we transferred the herpes simplex virus thymidine kinase gene (TK) into mouse Ltk/sup -/ cells. Independent TK/sup +/ clones (transformants) and derivatives of each were tested for phenotypic expression and the presence and arrangement of TK sequences. Initially, transformants expressed viral TK unstably, with 10% of the cells in each generation losing both the TK/sup +/ phenotype and virally derived TK sequences. After a prolonged period in culture, stable subpopulations arose from which the TK/sup +/ phenotype and viral sequence were no longer lost at detectable frequency. Analysis of unstable cell populations indicated that individual viral deoxyribonucleic acid molecules were reduced in size, but were linked to other deoxyribonucleic acid to form molecules large enough to be precipitated in a Hirt fractionation. The authors term these molecules transgenomes. Analysis of independent unstable subclones derived from the primary transformants demonstrated that individual transgenomes could contain multiple copies of the viral TK sequences. Recipient cell lines frequently possesses more than one type of the transgenome and possibly multiple copies per cell of each type. Stable derivatives possesses only one of the transgenomes present in the unstable parent, and these sequences were associated with a recipient cell chromosome.
- Research Organization:
- Dept. of Biology, The Johns Hopkins Univ., Baltimore, MD 21218
- OSTI ID:
- 5941798
- Journal Information:
- Mol. Cell. Biol.; (United States), Vol. 1:2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ANIMAL CELLS
GENOME MUTATIONS
HERPES SIMPLEX
DNA-CLONING
MOLECULAR BIOLOGY
PHOSPHOTRANSFERASES
GENE REGULATION
CELL CULTURES
CHROMOSOMES
CLONE CELLS
DNA
DNA SEQUENCING
FRACTIONATION
GENES
MESSENGER-RNA
ONCOGENIC VIRUSES
PHENOTYPE
RATS
THYMIDINE
TRANSCRIPTION
ANIMALS
AZINES
CLONING
DISEASES
ENZYMES
HETEROCYCLIC COMPOUNDS
INFECTIOUS DISEASES
MAMMALS
MICROORGANISMS
MUTATIONS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PHOSPHORUS-GROUP TRANSFERASES
PYRIMIDINES
RIBOSIDES
RNA
RODENTS
SEPARATION PROCESSES
SKIN DISEASES
STRUCTURAL CHEMICAL ANALYSIS
TRANSFERASES
VERTEBRATES
VIRAL DISEASES
VIRUSES
550400* - Genetics