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Isolation of Restriction Endonuclease Site Deletion Mutants of Herpes Simplex Virus

Institute of Virology, Church Street, Glasgow G11 5JR, U.K.

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 (TR_L or IR_L) 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 (TR_L or IR_L) have a measurable growth disadvantage in tissue culture.

Keywords: HSV-1, XbaI, deletion mutants

Received 6 December 1983; accepted 15 February 1984.