Determination of the recognition site for adenine-specific methylase of Shigella sonnei 47 by hydazinolysis of DNA, followed by separation of the purine oligonucleotides by thin-layer chromatography on DEAE-cellulose
Journal Article
·
· Biochemistry (Engl. Transl.); (United States)
OSTI ID:5722359
A method has been developed for the separation of oligopurine units according to length and composition by two-dimensional thin-layer chromatography on plates with DEAE-cellulose, permitting a comparative analysis of the content of various purine isopliths in DNA of different origin. In the case of the analysis of methylated DNA, the method permits a comparison of the substrate specificity of various enzymes of methylation of the adenine residues in DNA. In conjunction with enzymatic treatment of labeled methylated isopliths, the method permits determination of the methylatable sequence and in a number of cases an ascertainment of the recognition site for adenine-specific methylase as a whole. The proposed method was used to establish the fact that the methylase Ssol recognizes the sequence 5'...G-A-A-T-T-C...3' and methylates the adenine residue closest to its 5'-end.
- Research Organization:
- Scientific-Research Institute of Medical Enzymology, Moscow, USSR
- OSTI ID:
- 5722359
- Journal Information:
- Biochemistry (Engl. Transl.); (United States), Journal Name: Biochemistry (Engl. Transl.); (United States) Vol. 50:3; ISSN BIORA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADENINES
AMINES
ANTIMETABOLITES
AROMATICS
AZAARENES
BACTERIA
CHEMICAL REACTIONS
CHROMATOGRAPHY
DECOMPOSITION
DNA
DRUGS
ENZYMATIC HYDROLYSIS
HETEROCYCLIC COMPOUNDS
HYDROLYSIS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LABELLING
LYSIS
MEMBRANE PROTEINS
METHYLATION
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PROTEINS
PURINES
RADIOENZYMATIC ASSAY
RADIONUCLIDE KINETICS
RADIORECEPTOR ASSAY
RECEPTORS
SEPARATION PROCESSES
SHIGELLA
SOLVOLYSIS
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
UPTAKE
59 BASIC BIOLOGICAL SCIENCES
ADENINES
AMINES
ANTIMETABOLITES
AROMATICS
AZAARENES
BACTERIA
CHEMICAL REACTIONS
CHROMATOGRAPHY
DECOMPOSITION
DNA
DRUGS
ENZYMATIC HYDROLYSIS
HETEROCYCLIC COMPOUNDS
HYDROLYSIS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LABELLING
LYSIS
MEMBRANE PROTEINS
METHYLATION
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PROTEINS
PURINES
RADIOENZYMATIC ASSAY
RADIONUCLIDE KINETICS
RADIORECEPTOR ASSAY
RECEPTORS
SEPARATION PROCESSES
SHIGELLA
SOLVOLYSIS
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
UPTAKE