Isolation of restriction endonuclease site deletion mutants of herpes simplex virus.
We provide evidence that (i) variants lacking individual herpes simplex virus type 1 (HSV-1) XbaI sites can be selected following extensive XbaI treatment of the viral DNA and can be recombined to produce HSV-1 variants lacking two of the four sites normally found, (ii) all XbaI sites can be removed from a viable intertypic recombinant HSV genome, (iii) following XbaI treatment, different mutants with deletions (0.15 to 8.8 kb) in the long repeat ( TRL or IRL ) and long unique regions can be readily isolated, as well as mutants with novel XbaI sites, (iv) several mutants with deletions in one of the repeats ( TRL or IRL ) have a measurable growth disadvantage in tissue culture.
D001665:Binding Sites / Q000187:drug effects; D002872:Chromosome Deletion; D004262:DNA Restriction Enzymes / Q000378:metabolism* / Q000494:pharmacology; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D005814:Genes, Viral / Q000187:drug effects; D009154:Mutation; D009693:Nucleic Acid Hybridization; D012641:Selection, Genetic; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*
D004279:DNA, Viral; D004262:DNA Restriction Enzymes; C025580:endodeoxyribonuclease XBAI; D015252:Deoxyribonucleases, Type II Site-Specific