Methods and kits for identifying mutagenic agents and molecular mutations in DNA in mammalian cells

Patent · Tue Dec 20 04:00:00 UTC 1988

OSTI ID:5641202

Stanbrook, P J; Tischfield, J A

This patent describes a method of identifying a mutagenic agent which induces base substitution mutation in DNA in mammalian cells comprising the steps of: introducing a clone gene having a DNA sequence encoding a known selectable characteristic when in an unmutated form into nonreverting mammalian cells to produce transfectants of the mammalian cells, the untransfected nonreverting mammalian cells being nonselectable for the characteristics when cultured in a cell culture selection medium and the cloned gene having a base substitution at a specific site rendering the gene or its product nonselectable for the characteristic, whereupon the introduction of the cloned gene into the nonreverting mammalian cells renders the transfectants of the mammalian cells selectable for the characteristic when cultured in the cell culture medium following base substitution mutagenic reversion in the cloned gene at the specific site; exposing the transfected mammalian cells to a mutagenic agent for a sufficient amount of time to induce reversion by base substitution mutagenes in the cloned gene at the specific site; culturing the exposed transfected mammalian cells in the cell culture medium which selects for those transfected mammalian cells that are selectable for the characteristic as a result of base substitution mutagenic reversion in the cloned gene at the specific site; and identifying the mutagenic agent which has induced the base substitution mutation at the specific site in the cloned gene in the selected transfected mammalian cells.

• Assignee: Univ. of Cincinnati, Cincinnati, OH and Medical College of Georgia Research Institute, Augusta, GA
• Patent Number(s): US 4792520
• OSTI ID: 5641202
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
CELL CULTURES
CLONING
DNA
DNA BASE TRANSITIONS
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
EVALUATION
GENE MUTATIONS
GENES
GENETIC MAPPING
HYBRIDIZATION
MAMMALS
MAPPING
MOLECULAR BIOLOGY
MUTAGENESIS
MUTAGENS
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES