Expression of a cloned denV gene of bacteriophage T4 in Escherichia coli.
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 our earlier results [Valerie, K., Henderson, E. E. & de Riel, J. K. (1984) Nucleic Acids Res. 12, 8085-8096] that the denV gene is located at 64 kilobases on the T4 map. Phage T2 (denV-) did not hybridize to a denV-specific probe.
Valerie
K
K
Henderson
E E
EE
de Riel
J K
JK
D003001:Cloning, Molecular; D004260:DNA Repair; D004262:DNA Restriction Enzymes; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000235:genetics* / Q000528:radiation effects; D005796:Genes; D005814:Genes, Viral; D005822:Genetic Vectors; D009154:Mutation; D010957:Plasmids; D013604:T-Phages / Q000235:genetics* / Q000528:radiation effects; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000235:genetics*
D014764:Viral Proteins; D004262:DNA Restriction Enzymes