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Expression of a cloned denV gene of bacteriophage T4 in Escherichia coli

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
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A 713-base-pair Hae III fragment from bacteriophage T4 encompassing the denV gene with its preceding promoter has been cloned in a pBR322-derived positive-selection vector and introduced into a variety of DNA repair-deficient uvr and rec and uvr,rec Escherichia coli strains. The denV gene was found to be expressed, probably from its own promoter, causing pyrimidine dimer incision-deficient uvrA, uvrB, uvrC strains to be rescued by the denV gene. A uvrD (DNA helicase II) strain was also complemented, but to a lesser extent. A wild-type strain did not seem to be affected at the UV doses tested. Surprisingly, all recA, recB, and recC strains tested also showed an increased UV resistance, perhaps by reinforcement of the intact uvr system in these strains. Complementation of denV- T4 strains and host-cell reactivation of lambda phage was also observed in denV+ E. coli strains. Equilibrium sedimentation showed that DNA repair synthesis occurred in a UV-irradiated uvrA E. coli strain carrying the cloned denV gene. Southern blotting confirmed earlier results that the denV gene is located at 64 kilobases on the T4 map. Phage T2 (denV-) did not hybridize to a denV-specific probe.

Research Organization:
Fels Research Institute, Philadelphia, PA
OSTI ID:
5016934
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 14; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

560131* -- Radiation Effects on Microorganisms-- Basic Studies-- (-1987)
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
BACTERIA
BACTERIOPHAGES
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CONSTITUENTS
CLONING
DNA REPAIR
DOSE-RESPONSE RELATIONSHIPS
DOSES
ELECTROMAGNETIC RADIATION
ESCHERICHIA COLI
GENES
GENETIC EFFECTS
GENETIC RADIATION EFFECTS
HOST-CELL REACTIVATION
MICROORGANISMS
MUTATIONS
PARASITES
PLASMIDS
PYRIMIDINE DIMERS
RADIATION DOSES
RADIATION EFFECTS
RADIATIONS
RECOVERY
REPAIR
ULTRAVIOLET RADIATION
VIRUSES