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  • 1
    ISSN: 1432-072X
    Keywords: Archaea ; Methanogen ; Methanococcus deltae ; Flagella ; Glycosylation inhibition ; Bacitracin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methanococcus deltae is an irregularly-shaped coccoid methanogen which possesses peritrichously arranged flagella. The flagella are composed of 2 flagellins of M r=27000 and 32000 and possess a carbohydrate component as determined by thymol-sulfuric acid staining of SDS-PAGE. Cell growth was sensitive to bacitracin at levels near 10 μg/ml. Growth in the presence of 5μg/ml bacitracin resulted in the appearance of presumably hypoglycosylated flagellins as detected by Western immunoblotting. Continual passage of cells from 5 μg/ml bacitracin through 10, 25, and 50 μg/ml bacitracin resulted in cells capable of growth at 100 μg/ml bacitracin. Western immunoblot analysis revealed that cells grown in the highest concentration of bacitracin no longer possessed native flagellins but only the hypoglycosylated forms. Electron microscopy corroborated the absence of normal flagella. These studies suggest that a bacitracin-sensitive dolichol-diphosphate carrier is responsible for attachment of at least a large proportion of the carbonhydrate content of the flagellins, and that a minimum amount of glycosylation is essential for normal flagellum assembly.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The highly conserved nature of the 5′-termini of all archaeal flagellin genes was exploited by polymerase chain reaction (PCR) techniques to amplify the sequence of a portion of a flagellin gene family from the archaeon Methanococcus vannielii. Subsequent inverse PCR experiments generated fragments that permitted the sequencing of a total of three flagellin genes, which, by comparison with flagellin genes that have been sequenced, from other archaea appear to be equivalent to flaB1, flaB2, and flaB3 of M. voltae. Analysis of purified M. vannielii flagellar filaments by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed two major flagellins (Mr= 30 800 and 28 600), whose N-terminal sequences identified them as the products of the flaB1 and flaB2 genes, respectively. The gene product of flaB3 could not be detected in flagellar filaments by SDS-PAGE. The protein sequence data, coupled with the DNA sequences, demonstrated that both FlaB1 and FlaB2 flagellins are translated with a 12-amino acid signal peptide which is absent from the mature protein incorporated into the flagellar filament. These data suggest that archaeal flagellin export differs significantly from that of bacterial flagellins.
    Type of Medium: Electronic Resource
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