FIG 7.

SDS-PAGE and Western blotting of PDO-GFP fusions in E. coli BL21(DE3). The cells were suspended in 50 mM Tris-HCl buffer (pH 8.0) and disrupted; the lysate was centrifuged at 12,000 × g for 10 min to separate supernatant and cell debris, which was resuspended with the same buffer to the same volume. (A) SDS-PAGE. (B) Western blotting. Anti-GFP polyclonal antibody was used as the first antibody. Lanes 1 to 3, cell lysate (597.7 μg of protein ml⁻¹), supernatant (560.1 μg ml⁻¹), and debris resuspension (15.33 μg ml⁻¹), respectively, of E. coli BL21(DE3)(pBBR2-Cppdo2-gfp). Lanes 4 to 6, cell lysate (552.8 μg ml⁻¹), supernatant (533.4 μg ml⁻¹), and debris resuspension (12.2 μg ml⁻¹), respectively, of E. coli BL21(DE3)(pBBR2-Papdo2-gfp). Lanes 7 to 9, cell lysate (528.5 μg ml⁻¹), supernatant (504 μg ml⁻¹) and debris resuspension (21.8 μg ml⁻¹) of E. coli BL21(DE3)(pBBR2-mhpT-gfp). The sample volume was 15 μl for each well.