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Characterization of Aeromonas salmonicida variants with altered cell surfaces and their use in studying surface protein assembly

Abstract

Aeromonas salmonicida variants were characterized for alterations in their cell surface structure and used to examine reconstitution of the surface protein layer (A-layer). Variants lacking outer membrane O-polysaccharide were devoid of A-layer and excreted stainable floret-like material of the surface protein (A-protein). One variant, showing partial loss of O-polysaccharide, was associated with a disrupted A-layer and excretion of some A-protein. Variants lacking A-protein but possessing O-polysaccharide rapidly absorbed and concentrated sufficient excreted A-protein at the cell surface to coat the cells with a single confluent layer. Although differences in electrophoretic mobilities of A-proteins and O-polysaccharides from “typical” and “atypical” strains were evident, the different A-proteins and A-protein-deficient variants were interchangeable for reconstitution of a surface protein layer. No association of A-protein with cell surfaces of unrelated gram-negative bacteria was observed.

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Abbreviations

A-layer:

additional surface protein layer

A-protein:

surface protein

Ast:

Aeromonas salmonicida “typical”

Asa:

Aeromonas salmonicida “atypical”

A- :

phenotypically A-protein-negative variant

O- :

phenotypically O-polysaccharide-negative variant

Owk :

phenotypically O-polysaccharide weak variant

BHI:

brain heart infusion

SDS-PAGE:

sodium dodecylsulfate-polyacrylamide gel electrophoresis

TEM:

transmission electron microscopy

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Griffiths, S.G., Lynch, W.H. Characterization of Aeromonas salmonicida variants with altered cell surfaces and their use in studying surface protein assembly. Arch. Microbiol. 154, 308–312 (1990). https://doi.org/10.1007/BF00248973

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Key words

  • “typical” and “atypical” Aeromonas salmonicida variants
  • A-layer
  • O-polysaccharide
  • A-protein reconstitution