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Expression and cellular localization of the BBB26-FLAG and BBB27-FLAG proteins

figure

posted on 2011-12-31, 12:37 authored by Mollie W Jewett, Rebecca Byram, Aaron Bestor, Kit Tilly, Kevin Lawrence, Mary N Burtnick, Frank Gherardini, Patricia A Rosa

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Taken from "Genetic basis for retention of a critical virulence plasmid of "

Molecular Microbiology 2007;66(4):975-990.

Published online 05 Nov 2007

PMCID:PMC2229028.

A. Proteins lysates from (Ec) or B31-A34 (Bb) harbouring either pBSV2ex -FLAG or pBSV2ex -FLAG were separated by SDS-PAGE and analysed by immunoblot with anti-FLAG antibodies. The mobilities of size standards (molecular weights in kDa) are indicated to the left of the figure. B. Protein lysates from clone A34 harbouring either pBSV2ex -FLAG or pBSV2ex -FLAG were harvested and separated into soluble and membrane fractions by ultracentrifugation. Protein fractions from equivalent numbers of spirochetes were subjected to SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), OppAIV (inner membrane), OspB (outer membrane) and SodA (cytoplasmic) antisera. Representative results for the localization of the OppAIV, OspB and SodA proteins from clone A34 harbouring either pBSV2ex -FLAG or pBSV2ex -FLAG are shown. L, total cell lysate; S, soluble protein fraction; M, membrane protein fraction. C. Equal numbers of whole or 0.1% SDS-treated (+0.1% SDS) cells of clone A34 harbouring either pBSV2ex -FLAG or pBSV2ex -FLAG were incubated with different concentrations (μg ml) of proteinase K (PK). Lysates of PK-treated bacteria were separated by SDS-PAGE and analysed by immunoblot with FLAG (BBB26 and BBB27), flagellin (FlaB, periplasmic marker), OppAIV (inner membrane marker) and OspB (surface exposed, outer membrane marker) antisera. Representative results for the proteinase K sensitivity of the FlaB, OppAIV and OspB proteins from clone A34 harbouring either pBSV2ex -FLAG or pBSV2ex -FLAG are shown.