Mutagenesis of the lac promoter region in M13 mp10 phage DNA by 4'-hydroxymethyl-4,5',8-trimethylpsoralen
Double-stranded M13 phage DNA (M13 mp10 replicative form) was photoreacted with 4'-hydroxymethyl-4,5',8-trimethylpsoralen, using light of wavelength greater than 320 nm or greater than 390 nm to generate predominantly crosslinks or monoadducts, respectively. The damaged DNAs were scored for inactivation and mutagenesis after transfection into Escherichia coli. The appearance of light-blue or colorless plaques on indicator medium showed that mutation had occurred in the lac insert of the viral DNA. A comparison of the consequences of the two phototreatments with psoralen supports the idea that crosslinks are both more lethal and more mutagenic than monoadducts. Numerous mutant clones partially or totally deficient in beta-galactosidase were plaque-purified and amplified. The viral DNA of each clone was sequenced by the dideoxy chain-terminating procedure. All of the observed base-pair changes were mapped to the lac promoter region and consisted of 3 transition, 14 transversion, and 6 single base-pair frame-shift mutations. The predominant mutation was a T.A----G.C transversion.
- Research Organization:
- Univ. of Liege, Belgium
- OSTI ID:
- 6255734
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 21
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
CROSS-LINKING
RADIOINDUCTION
DNA
PHOTOCHEMISTRY
DNA ADDUCTS
BACTERIOPHAGES
DNA SEQUENCING
ESCHERICHIA COLI
GENE MUTATIONS
NEAR ULTRAVIOLET RADIATION
PSORALEN
ADDUCTS
ANTICOAGULANTS
BACTERIA
CHEMICAL REACTIONS
CHEMISTRY
COUMARINS
DRUGS
ELECTROMAGNETIC RADIATION
HEMATOLOGIC AGENTS
HETEROCYCLIC COMPOUNDS
MICROORGANISMS
MUTATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PARASITES
POLYMERIZATION
RADIATIONS
STRUCTURAL CHEMICAL ANALYSIS
ULTRAVIOLET RADIATION
VIRUSES
560111* - Radiation Effects on Biochemicals- In Vitro- (-1987)
550400 - Genetics