FIG. 5.

Phenotype of a kin28^T162A strain. (A) kin28Δ cells containing a URA3-marked wild-type (WT) KIN28 plasmid (pJK1) and TRP1-marked plasmids containing either KIN28 (pGK13), kin28^T162A (pGK36), kin28-ts16 (pGK33), kin28^D147N (pMS454), or kin28-ts^T162A (pJK25) were plated on FOA-containing plates to select for loss of the URA3 marked plasmid. (B) kin28Δ cells containing KIN28 (YJK1869, YJK1767, and YJK1768) or kin28^T162A (YJK1870, YJK1755, and YJK1756) and either an empty vector (YCplac22), kin28^D147N on a low-copy-number plasmid (pMS454), or kin28^D147N on a high-copy-number plasmid (pMS456) were plated on CM-Trp and grown at 37°C.