A nonradioactive DNA methyltransferase assay adaptable to high-throughput screening.
Anal Biochem
; 340(2): 336-40, 2005 May 15.
Article
de En
| MEDLINE
| ID: mdl-15840507
ABSTRACT
We have developed a nonradioactive assay method for DNA methyltransferases based on the ability to protect substrate DNA from restriction. DNA immobilized to a microplate well was treated sequentially with methyltransferase and an appropriate endonuclease. The amount of methylated DNA product is reflected by a proportional decrease in endonuclease cleavage, which is in turn reflected by increased retention of the end-labeled affinity probe. A single universal substrate was designed to assay multiple methyltransferases including those that do not have a cognate endonuclease. The methodology developed is suited to screen a large number of compounds for inhibitors of various methyltransferases.
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Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
DNA modification methylases
Type d'étude:
Diagnostic_studies
/
Screening_studies
Langue:
En
Journal:
Anal Biochem
Année:
2005
Type de document:
Article
Pays d'affiliation:
États-Unis d'Amérique