FIG. 3.

Deletion of PKC1 requires osmotic stabilization, is temperature sensitive, and is complemented by reintroduction of the PKC1 gene. (A) Strains with PKC1 deletions in two genetic backgrounds, KN99a or cku80Δ (KN99α), were streaked onto YPD plates with or without 1 M sorbitol and grown at 25, 30, or 37°C for 5 days. The pkc1Δ strains failed to grow in the absence of sorbitol at any temperature and in the presence of sorbitol at 37°C. The parental strains (KN99a or cku80Δ in the KN99α background) were also plated as controls. wt, wild type. (B) Temperature sensitivity of pkc1Δ is rescued in the complemented strain. The pkc1Δ strain in the KN99a background (KN99a) and the complemented pkc1Δ::PKC1 strain were grown overnight in YPD plus 1 M sorbitol and then diluted to an OD₆₅₀ of 1.0. Tenfold serial dilutions were plated onto YPD plates containing 1 M sorbitol and grown for 5 days at 30, 37, or 39°C.