Figure 5.

Electrophoretic mobility of Pah1 from spo7Δ cells expressing Spo7 with LLI mutations. The spo7Δ mutant (GHY67) was transformed with pGH443 and its derivatives for expression of the WT and mutant forms of Spo7-Myc. The yeast transformants were grown at 30 °C in SC-Leu medium to the mid-logarithmic phase. Cell extracts were prepared and subjected to SDS-PAGE (40 µg protein) using an 8% polyacrylamide gel. A, the proteins resolved in the polyacrylamide gel were transferred to a PVDF membrane and probed with anti-Pah1 antibody. The positions of Pah1 and molecular mass standards are indicated. The white dashed line is a guide to show the range in the electrophoretic mobility of Pah1 from the spo7Δ cells and those expressing WT Spo7. B, the signal intensities of Pah1 along its migration in the region between the dashed white lines in panel A were measured using the line graph function of ImageQuant software. The densitogram of Pah1 from the expression of mutant Spo7 (black line) was compared with those from the presence (green line) and absence (red line) of WT Spo7. The vector control and WT lines are included with each mutant protein for comparison. The data shown are representative of three separate experiments.