Journal: Applied and Environmental Microbiology

Article Title: Changing a Single Amino Acid in Clostridium perfringens β-Toxin Affects the Efficiency of Heterologous Secretion by Bacillus subtilis

doi: 10.1128/aem.02356-06

Figure Lengend Snippet: FIG. 4. Degradation of -toxin and -toxoid by spent B. subtilis supernatants. Lactococcus lactis-produced -toxin and -toxoid cul- tures were incubated with spent supernatants of stationary-phase cul- tures of B. subtilis strains 168 and WB800. As a control, TY medium was used. (A) Typical Coomassie brilliant blue-stained polyacrylamide gel showing results of the degradation assay. The incubation time (T) in minutes is indicated. Left lanes show -toxoid and -toxin exposed to strain 168 culture supernatant; right lanes show -toxoid and -toxin exposed to strain WB800 culture supernatant. M, protein marker, 35-kDa band. (B) The amount of -toxin/-toxoid measured after 10 min was set to 100%. The remaining amounts of -toxoid and -toxin after 1 h were determined and plotted. Experiments were performed in duplicate; error bars depict standard errors.

Article Snippet: The concentrations of -toxin and -toxoid were determined by Coomassie brilliant blue staining followed by densitometric scanning (Bio-Rad GS-800 scanner) and analysis with Quantity One software (Bio-Rad, Hercules, CA).

Techniques: Produced, Incubation, Control, Staining, Degradation Assay, Marker

Journal: Applied and Environmental Microbiology

Article Title: Changing a Single Amino Acid in Clostridium perfringens β-Toxin Affects the Efficiency of Heterologous Secretion by Bacillus subtilis

doi: 10.1128/aem.02356-06

Figure Lengend Snippet: FIG. 5. Secretion and secretion stress of -toxin, -toxoid, and intermediate mutants. (A) Coomassie brilliant blue-stained 12% SDS- polyacrylamide gel containing 10 concentrated supernatant of B. subtilis strain NZ8900 1.5 h after induction of the inducible tox plasmids. (B) The average PhtrA-GFP expression per cell in arbitrary units, measured 1.5 h after induction, is shown. Experiments were performed in duplicate; error bars depict standard errors. DDK, WT -toxin (D54DK); AAA, -toxoid (A54AA); all other intermediate mutants (mut) are likewise indicated.

Article Snippet: The concentrations of -toxin and -toxoid were determined by Coomassie brilliant blue staining followed by densitometric scanning (Bio-Rad GS-800 scanner) and analysis with Quantity One software (Bio-Rad, Hercules, CA).

Techniques: Staining, Expressing