Figure 2.

Detection of the full-length win1⁺ gene product. (A) Strategy for constructing a strain carrying tagged Win1 (ED1423) by homologous recombination. The win1⁺ gene in the haploid wild-type strain was disrupted by the S. pombe ura4⁺ gene to generate a win1Δ strain (ED1370). Then the ura4⁺ marker of the deletion strain was replaced by a sequence encoding the tagged carboxyl-terminal domain of Win1, using 5-fluoro-orotic acid selection (Grimm et al., 1998). The shaded box indicates the kinase domain. (B) Western blot of S. pombe cell extract from ED1423. Anti-HA antibody (12CA5) was used as the primary antibody.