Herpes simplex virus type 2 differs from type 1 virus in the sites it affects and in certain biological and serological properties (Schneweis, 1962; Plummer, 1964; Hutfield, 1967; Figueroa & Rawls, 1969). Temperature-sensitive mutants of type 1 virus have been isolated and characterized (Subak-Sharpe, 1969; Schaffer 1971; Brown, Ritchie & Subak-Sharpe, 1971). This report describes the isolation and preliminary classification of temperature-sensitive mutants of type 2 virus.

BHK 21 clone 13 cells (Macpherson & Stoker, 1962) and Eagles medium with 10% (v/v) calf serum and 10% (v/v) tryptose phosphate broth were used throughout. Virus was titrated in suspended cells by the method described by Russell (1962); titrations at 38° were incubated for 2 days and those at 31° for 3 days. For assays under agar — which were always carried out at 31° — virus was adsorbed to cell monolayers in 60 mm. diameter Petri plates for 1 hr before addition of medium containing 0.9% (w/v) Difco Bacto agar; plates were stained with neutral red after 6 days.


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