SDS-PAGE and immunoblot analysis showing the effects of recombinant plasmid pLPSα and mutant plasmid pLPSα395. After growth in YEC, with or without seed extract, washed bacteria were processed for SDS-PAGE. (A) Strains CE3 (lanes 1), CE395 (lanes 2), and CE395α395 (lanes 3) grown in YEC without seed extract are compared. (B) Strains carrying pLPSα (CE3α and CE395α) are included; the effects of seed extracts are shown. The analyzed strains were CE3 (lanes 1 and 2), CE395 (lanes 3 and 4), CE3α (lanes 5 and 6), CE395α (lanes 7 and 8), and CE395α395 (lanes 9 and 10). In lanes 2, 4, 6, 8, and 10 of panel B, the strains had been grown in the presence of seed extract, which is responsible for the streaking above LPS I in the silver-stained gel. In each panel the image on the left is of gel lanes stained with silver. (In panel B the gel was stained after being elecroblotted, resulting in relatively less-intense staining of LPS II bands.) On the right in each panel is the blot of these lanes reacted with monoclonal antibody JIM28.