FIG. 2.
(A) Agarose plate assays for Prt and Cel activities of E. carotovora subsp. carotovora AC5047 (a and c) and its RsmC− mutant (AC5050; b and d) carrying the cloning vector, pCL1920 (column 1) or the RsmC+ plasmid, pAKC975 (column 2). Bacteria were grown at 28°C in minimal salts medium plus sucrose and spectinomycin and harvested at an A600 of 2.5. Culture supernatants (10 μl) were added to each well. (B) Northern blot analysis of pel-1, peh-1, celV, hrpNEcc, and rsmB transcripts produced by E. carotovora subsp. carotovora AC5047 (RsmC+) and AC5050 (RsmC−) carrying the cloning vector pCL1920 or the RsmC+ plasmid pAKC975. Lane 1, AC5047 carrying pCL1920; lane 2, AC5047 carrying pAKC975; lane 3, AC5050 carrying pCL1920; lane 4, AC5050 carrying pAKC975. Total RNAs were isolated from bacteria grown at 28°C in minimal salts medium plus sucrose and spectinomycin to an A600 of 2.0. Each lane contained 10 μg of total RNA. (C) Northern blot analysis of pel-1, peh-1, celV, and hrpNEcc mRNA produced by E. carotovora subsp. carotovora Ecc71 (RsmA+ RsmC+), AC5053 (RsmA+ RsmC−), AC5071 (RsmA− RsmC+), and AC5054 (RsmA− RsmC−) carrying the cloning vector pCL1920Gmr or the rsmB+ plasmid pAKC1004Gmr. Lane 1, Ecc71/pCL1920Gmr; lane 2, Ecc71/pAKC1004Gmr; lane 3, AC5053/pCL1920Gmr; lane 4, AC5053/pAKC1004Gmr; lane 5, AC5071/pCL1920Gmr; lane 6, AC5071/pAKC1004Gmr; lane 7, AC5054/pCL1920Gmr; lane 8, AC5054/pAKC1004Gmr. Total RNAs were isolated from bacteria grown at 28°C in minimal salts medium plus sucrose and gentamicin to an A600 of 2.0. Each lane contained 10 μg of total RNA.