FIG. 2.

Immunoblots of GFP-tagged SpoIIE derivatives during vegetative growth and sporulation with anti-GFP antibodies. (A) SDS-polyacrylamide gel electrophoresis Western blot of lysates prepared from vegetative cells uninduced (SB220) or induced with 0.5 mM IPTG (SB220 [wild-type SpoIIE-GFP], SB414 [SpoIIE^S361F-GFP], KC507 [SpoIIE^Q483A-GFP], KC552 [SpoIIE^D686A-GFP], KC506 [MalF-SpoIIE-GFP], SB221 [SpoIIE^ΔregII-GFP], KC542 [SpoIIE^ΔregI-GFP]) and probed with anti-GFP antibodies (upper panel) and anti-σ^A antibodies (lower panel). (B) SDS-polyacrylamide gel electrophoresis Western blot of lysates prepared from cells harvested 2 h after the start of sporulation (KC544 [wild-type SpoIIE-GFP], KC545 [SpoIIE^S361F-GFP], KC546 [SpoIIE^Q483A-GFP], KC547 [SpoIIE^D746A-GFP], KC554 [SpoIIE^D686A-GFP], KC538 [MalF-SpoIIE-GFP], KC548 [SpoIIE^ΔregII-GFP], and KC549 [SpoIIE^ΔregI-GFP]) and probed with anti-GFP antibodies (upper panel) and anti-σ^A antibodies (lower panel). In each upper panel the top asterisk indicates the position on the gel of full-length SpoIIE-GFP, the middle asterisk marks SpoIIEΔregI-GFP, and the bottom asterisk marks SpoIIEΔregII-GFP. WT, wild type. Numbers at left represent molecular masses in kilodaltons.