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. Author manuscript; available in PMC: 2011 Nov 1.
Published in final edited form as: Mol Microbiol. 2010 Sep 14;78(3):710–719. doi: 10.1111/j.1365-2958.2010.07362.x

Figure 2. A. Representatives of DSP crosslinking of S2168 varients in vivo.

Figure 2

Whole cells carrying prophage λQ21Δ(SRRzRz1)21 only (lane 1) or with each indicated plasmid were induced and, treated with the cross-linker DSP, and analyzed by SDS-PAGE and western blotting, as described in the Materials and Methods. Plasmid carried in each sample: pS2168 (lane 2), pirsS2168* (lane 3), lane 4–9, derivatives of pS2168, encoding single mutation of S2168 as indicated on top of each lane. Numbers at the right side of each lane indicate the crosslinked oligomers.

B. Blue-native PAGE separation of S2168 and its variants.

Crude membranes from cells carrying prophage λQ21Δ(SRRzRz1)21 only (lane 1) or with derivatives of plasmid pS2168 expressing S2168 (lane 2) or its mutants (lanes 3–6) were analyzed by BN PAGE and immunoblotting, as described in Materials and Methods. The position of oligomeric states of S2168 and its variants were estimated by their apparent molecular weights with detergent micelles. The position of BN PAGE standards are indicated: 242 kDa = B-phycoerythrin; 146 kDa = lactate dehydrogenase; 66 kDa = bovine serum albumin; 20 kDa = soybean trypsin inhibitor.