Journal:

Article Title: Cloning of an Intracellular Poly[ d (-)-3-Hydroxybutyrate] Depolymerase Gene from Ralstonia eutropha H16 and Characterization of the Gene Product

doi: 10.1128/JB.183.1.94-100.2001

Figure Lengend Snippet: Detection of PhaZ in R. eutropha H16. SDS-polyacrylamide gel electrophoresis of whole cells and PHB granules from R. eutropha was carried out. The gels were stained with Coomassie brilliant blue R-250 (A) or immunostained (Western blot) (B). Lanes: a-0, whole cells grown for 2 days in nitrogen-rich medium (91 μg of protein); a-1, whole cells after 1 day of PHB synthesis (86 μg of protein, 21 μg of PHB); a-3, whole cells after 3 days of PHB synthesis (88 μg of protein, 73 μg of PHB); b-1, PHB granules after 1 day of PHB synthesis (19 μg of protein, 160 μg of PHB); b-3, PHB granules after 3 days of PHB synthesis (20 μg of protein, 200 μg of PHB); c-1, supernatant fraction of cells after 1 day of PHB synthesis (91 μg of protein); c-3, supernatant fraction of cells after 3 days of PHB synthesis (57 μg of protein); Ni, purified His-tagged depolymerase (0.1 μg of protein); M, molecular mass markers. Sizes are indicated in kilodaltons.

Article Snippet: After metal chelating purification, the protein was finally purified by a continuous elution sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (Prep cell model 491; Bio-Rad).

Techniques: Polyacrylamide Gel Electrophoresis, Staining, Western Blot, Purification