Journal: Biomolecules
Article Title: The Passage of Chaperonins to Extracellular Locations in Legionella pneumophila Requires a Functional Dot/Icm System
doi: 10.3390/biom15010091
Figure Lengend Snippet: Immunoblots of SDS-PAGE-resolved proteins from whole bacterial cells treated with trypsin (250 µg trypsin/10 9 bacteria), and immunostained with an HtpB-specific monoclonal antibody and alkaline phosphatase-conjugated rabbit anti-mouse IgG. ( A ) Band patterns produced in the wild-type strain Lp02 and its dotA − derivative JV309. Trypsin-accessible HtpB is evidenced by the immunolabeled degradation products of <60 kDa. The presence of trypsin-accessible HtpB was recovered in the trans -complemented dotA − mutant ( dotA − + C). ( B , C ) Band patterns showing the lack of trypsin-accessible HtpB in two different dotB mutants: dotB − (with a loss-of-function point mutation), and Δ dotB (with a deletion of the dotB gene). A recovery of trypsin-accessible HtpB was observed in trans -complemented dotB mutants ( dotB + C and Δ dotB + C), but not in the mock-complemented mutants carrying an empty vector ( dotB + V and Δ dotB + V). Abbreviations for all panels: “C” = control, trypsin-free sample, “T” = trypsin-treated sample. Arrows at the far left indicate the position of three of the broad-range pre-stained protein size markers (New England Biolabs, Whitby, ON, Canada) corresponding to (from top to bottom) 62, 47 and 37.5 kDa.
Article Snippet: αMouseAP , Commercial rabbit purified immunoglobulin G conjugated to alkaline phosphatase. Diluted 1:5000 for WB , Cedarlane Labs 5.
Techniques: Western Blot, SDS Page, Bacteria, Produced, Immunolabeling, Mutagenesis, Plasmid Preparation, Control, Staining