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. 2006 Oct 27;189(1):207–219. doi: 10.1128/JB.00950-06

FIG. 3.

FIG. 3.

Subcellular localization of P. putida DOT-T1E(PP3005). (A) Hydrophobicity (Kyte-Doolittle) plot of the PP3005 protein showing a potential transmembrane segment (window size, 19; peaks with scores greater than 1.8 indicate putative transmembrane α-helices). (B) Schematic view of two different PP3005′-′phoA fusions under the control of the Plac promoter in plasmids pB3005AP-S and pB3005AP-L, indicating the expression of alkaline phosphatase activity. The numbers indicate the positions of the first and last amino acid residues of the proposed transmembrane domain. (C) Immunoblot detection of the PP3005′-′PhoA fusions in P. putida DOT-T1E cell fractions. Protein samples were subjected to electrophoresis (12.5% [wt/vol] SDS-PAGE), followed by Western blotting with an anti-PhoA antibody. Lanes 1 through 4, P. putida DOT-T1E(pB3005AP-S); lanes 5 through 8, P. putida DOT-T1E(pP3005AP-L). Lanes 1 and 5, whole-cell lysate (W); lanes 2 and 6, periplasmic fraction (P); lanes 3 and 7, cytoplasmic fraction (C); and lanes 4 and 8, membrane protein fraction (M).