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Cells. BHK-21 clone 13 cells (10) were grown in Eagle medium supplemented with 10% calf serum. Virus. The type 1 parent strain 17 and temperature-sensitive (ts) mutants used in this study, tsA,-B,-D,-E,-H,-K, and-S, have previously been described (3, 11). Syn+ forns tsH syn and tsS syn were isolated by M. Brown (unpublished data). The type 2 parent strain, HG52, and mutants tsl,-5,-6, and-11 were described by Timbury (18) and Halliburton and Timbury (8). tsll PAA' is a phosphonoacetic acid-resistant derivative of tsll (Timbury, unpublished data). The isolation and genome structures of the recombinants used in this study are summarized in Table 1 and Fig. 1. The previously undescribed recombinants B x 5 (7), B x 5 (10), B x 6 (17), 17+ x llr (1), and F x 5 (12) were generated and plaque purified by the 624

Twenty-three complementation groups of herpes simplex virus type 1 (HSV-1) and 20 of HSV-2 were identified by qualitative and quantitative complementation analysis from among 43 temperature-sensitive (ts) mutants of HSV-1 and 29 ts mutants of HSV-2 which had been isolated independently in 10 laboratories.

Recombinants between temperature-sensitive mutants of herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) were constructed. Using restriction endonucleases, we analyzed the genome composition of 17 intertypic recombinants and detected crossovers in every region of the genome. The virion DNA of one recombinant appeared to be largely "frozen" in two of the four possible genome arrangements of HSV. Knowledge of the genome structures of recombinants enabled us to physically map immediate early polypeptides. We present evidence that the immediate early polypeptide Vmw IE 110 of HSV-1 and its functionally equivalent polypeptide, Vmw IE 118, of HSV-2 may map in the repetitive sequences bounding the long unique region of HSV. MATERUILS AND METHODS Cells. BHK-21 clone 13 (C13) cells (16) were used in all experiments except in the isolation of certain HSV-1/HSV-2 recombinants, in which Vero cells were used. BHK-21 (C13) cells were cultivated in Eagle medium containing twice the concentration of vitamins and amino acids, 10% (vol/vol) tryptose phosphate broth, and 10% (vol/vol) calf serum. Vero cells were grown in this medium except that 10% (vol/vol) fetal calf serum (Gibco-Biocult) was used instead of calf serum. 499

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