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Three Mutants of Herpes Simplex Virus Type 2: One Lacking the Genes US10, US11 and US12 and Two in which R_S Has Been Extended by 6 kb to 0·91 Map Units with Loss of U_S Sequences between 0·94 and the U_S/TR_S Junction

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

In the process of generating restriction endonuclease site deletion mutants, we have isolated and characterized three mutants of herpes simplex virus type 2 (HSV-2), strain HG52, with large genomic deletions in U_S and TR_S. The deleted sequences (7·5 kb) extend from 0·94 map coordinates (m.c.) to 0·99 m.c. and are presumed to contain the HSV-2 gene equivalents of US10, 11 and 12, one copy of immediate early (IE) gene 3 and one copy of an origin of replication (ORI_S). One of the mutants (HG52X163X12) has a simple deletion whereas in the two others (HG52X163X14 and HG52X163X21) the deleted sequences have been replaced by inverted duplications of U_S/IR_S sequences between 0·83 and 0·91 m.c. such that the molecules have short region inverted repeats extended by 6 kb on either side. All three are viable, stable and grow in tissue culture indicating that the polypeptides coded by the HSV-2 genes equivalent to US10, 11 and 12 are non-essential for lytic growth in BHK21/C13 cells. In addition the lack of one copy of the HSV-2 equivalent of IE gene 3 and ORI_S in HG52X163X12 shows that only one copy of each suffices for viability. However the temperature restriction of the mutants at 38·5 °C suggests that one or more of the polypeptides coded by the deleted sequences may be required in conjunction with another polypeptide(s) for viral growth or stability at the higher temperature.

Received 28 May 1986; accepted 28 August 1986.

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