Method for identifying mutagenic agents which induce large, multilocus deletions in DNA
A method of identifying a mutagenic agent is described which includes a large, multilocus deletions in DNA in mammalian cells comprising: (i) exposing a class III heterozygous CHO cell line to a potential mutagenic agent under investigation, and allowing any mutation of the cell line to proceed, said cell line being characterized in that a restriction fragment length variation exists in on mutation it becomes resistant to 2,6-diaminopurine and in that the DNA sequence adjacent to the two alleles of the APRT gene such that the DNA sequence adjacent to one of the two alleles can be digested with the enzyme BclI but the DNA sequence variation adjacent to the other of the two alleles cannot be digested with BclI, (ii) isolating induced mutations of the cell line deficient in APRT function, (iii) isolating DNA from the induced mutants, (iv) digesting the isolated DNA with BclI enzyme to produce digested fragments including a 19 kb fragment and any 2 kb fragment, which fragments hybridize with the labeled probe derived from DNA fragment PDI, (v) separating any digested fragments, (vi) transferring the separated fragments of (v) to a solid support, (vii) hybridizing the supported separated fragments with a labeled probe derived from the clone DNA fragment PD 1, (viii) determining fragments having undergone loss of the 2 kb band identified by the probe, as an identification of parent mutants in which the loss occurred, and (ix) evaluating the mutating ability of the potential mutagenic agent.
- Assignee:
- IMS; EDB-93-110249
- Patent Number(s):
- US 5227289; A
- Application Number:
- PPN: US 7-524551
- OSTI ID:
- 7369141
- Resource Relation:
- Patent File Date: 17 May 1990
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
CHO CELLS
MUTATIONS
MUTAGEN SCREENING
DNA
DNA HYBRIDIZATION
DNA SEQUENCING
ENZYMES
GENES
GENETIC EFFECTS
MAMMALS
MUTAGENS
ANIMAL CELLS
ANIMALS
BIOLOGICAL EFFECTS
HYBRIDIZATION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
SCREENING
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology