6533b824fe1ef96bd12814d3
RESEARCH PRODUCT
A new assay for O6-alkylguanine-DNA-alkyltransferase to determine DNA repair capacities using lambda-phage DNA as substrate.
Franz OeschS. Kleinsubject
Cancer ResearchGuanineDNA RepairDNA repairMolecular Sequence DataBiologySubstrate Specificitychemistry.chemical_compoundO(6)-Methylguanine-DNA MethyltransferaseDNA Repair ProteinEscherichia coliHumansLymphocyteschemistry.chemical_classificationBase SequenceSubstrate (chemistry)General MedicineMethyltransferasesLambda phagebiology.organism_classificationBacteriophage lambdaIn vitroKineticsEnzymeBiochemistrychemistryDNA ViralBamHIDNAdescription
One O6-methylguanine (O6-meG) was introduced into each BamHI site of lambda-phage DNA as a substrate for the determination of the DNA repair protein O6-alkylguanine-DNA-alkyltransferase. A new assay using as the detection group 32P-labeled phosphate introduced at the 3' position of the modified nucleoside by incorporation of 32P-labeled TTP in the 3'-neighboring position proved highly sensitive: 10(-16) mol of the DNA lesion was still easily detectable. This DNA, which has greater than 1000 bp represents a good model for cellular DNA and was used as a substrate to measure the individual repair capacities for O6-meG in human lymphocytes of 20 healthy male and female donors. There were great inter-individual variations in the activity of this repair protein in man. The influences of age, sex or smoking behavior on the repair capacity of O6-meG were negligible.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1990-10-01 | Carcinogenesis |