Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Efficient synthesis of supercoiled M13 DNA molecule containing a site specifically placed psoralen adduct and its use as a substrate for DNA replication

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00409a013· OSTI ID:7130773
;

The authors report a simple method for the in vitro synthesis of large quantities of site specifically modified DNA. The protocol involves extension of an oligonucleotide primer annealed to M13 single-stranded DNA using part of the T4 DNA polymerase holoenzyme. The resulting nicked double-stranded circles are ligated and supercoiled in the same tube, producing good yields of form I DNA. When the oligonucleotide primer is chemically modified, the resultant product contains a site-specific lesion. In this study, they report the synthesis of an M13 mp19 form I DNA which contains a psoralen monoadduct or cross-link at the KpnI site. They demonstrate the utility of these modified substrates by assessing the ability of the bacteriophage T4 DNA replication complex to bypass the damage and show that the psoralen monoadduct poses a severe block to the holoenzyme when attached to the template strand.

Research Organization:
Univ. of Texas, Austin (USA)
OSTI ID:
7130773
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 27:9; ISSN BICHA
Country of Publication:
United States
Language:
English

Similar Records

Photochemistry of psoralen-DNA adducts, biological effects of psoralen-DNA adducts, applications of psoralen-DNA photochemistry
Technical Report · Mon Feb 29 23:00:00 EST 1988 · OSTI ID:5069947
The application of psoralens to the study of DNA structure, function and dynamics
Thesis/Dissertation · Sun Mar 31 23:00:00 EST 1991 · OSTI ID:10115059
Base-unpaired regions in supercoiled replicative form DNA of coliphage M13
Journal Article · Thu Aug 25 00:00:00 EDT 1977 · J. Biol. Chem.; (United States) · OSTI ID:5045902

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ADDUCTS
ANTICOAGULANTS
ATP
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOSYNTHESIS
CONFORMATIONAL CHANGES
COUMARINS
DAYS LIVING RADIOISOTOPES
DNA
DNA ADDUCTS
DNA REPLICATION
DRUGS
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
HEMATOLOGIC AGENTS
HETEROCYCLIC COMPOUNDS
IN VITRO
ISOTOPES
LIGHT NUCLEI
NEAR ULTRAVIOLET RADIATION
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PSORALEN
RADIATION EFFECTS
RADIATIONS
RADIOINDUCTION
RADIOISOTOPES
SYNTHESIS
ULTRAVIOLET RADIATION