Comparative studies of Yersinia pestis outer membrane isolation techniques and their potential use in plaque epidemiology.
In the present study three techniques for obtaining outer membrane enriched fractions from Yersinia pestis were evaluated. The techniques analysed were: differential solubilization of the cytoplasmic membrane with Sarkosyl or Triton X-100, and centrifugation in sucrose density gradients. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of outer membrane isolated by the different methods resulted in similar protein patterns. The measurement of NADH-dehydrogenase and succinate dehydrogenase (inner membrane enzymes) indicated that the outer membrane preparations obtained by the three methods were pure enough for analytical studies. In addition, preliminary evidences on the potential use of outer membrane proteins for the identification of geographic variants of Y. pestis wild isolates are presented.
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Centro de Pesquisas Aggeu Magalhães (Fundação Oswaldo Cruz), Pernambuco, Brazil.
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D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification*; D002462:Cell Membrane / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D009245:NADH Dehydrogenase / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols; D013385:Succinate Dehydrogenase / Q000378:metabolism; D015010:Yersinia pestis / Q000032:analysis*
D001425:Bacterial Outer Membrane Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; D013385:Succinate Dehydrogenase; D009245:NADH Dehydrogenase