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  • 1
    Electronic Resource
    Electronic Resource
    Expression, isolation and properties of Fur (ferric uptake regulation) protein ofEscherichia coli K 12 (1988)
    Springer
    BioMetals 1 (1988), S. 62-68 
    Details
    ISSN: 1572-8773
    Keywords: Fur repressor ; Iron regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The clonedfur (ferric uptake regulation) gene ofEscherichia coli K12 was ligated to an expression vector which was inducible with nalidixic acid. The Fur protein was isolated in a single step by immobilized metal-ion-affinity chromatography over zinc iminodiacetate agarose. The amino acid composition of the isolated protein agreed with that predicted from the gene sequence and indicated post-transcriptional removal of the N-terminal methionine residue. All four cysteines were shown to be present as thiols. Proteolysis with trypsin and chymotrypsin yielded large fragments identifiable on polyacrylamide gel electrophoresis. Various divalent metal ions were found by a nitrocellulose filter binding assay to effect non-specific interaction of the Fur dimer with DNA with a dissociation constant of 7 × 10−12 M. A much smaller value, 2.5 × 10−17 M, was measured by gel mobility retardation assay for binding of Fur to a DNA fragment containing the operator sequences of the aerobactin promoter.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01128019
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    IS1-mediated mobility of the aerobactin system of pColV-K30 in Escherichia coli (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 487-490 
    Details
    ISSN: 1617-4623
    Keywords: pColV ; Aerobactin ; Transposition ; IS1 ; Cointegration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genes determining the high affinity iron transport system mediated by the siderophore aerobactin are flanked in the enterobacterial plasmid pColV-K30 by inverted repeats of IS1 sequences, suggesting that the aerobactin genes are part of a transposon. To study this possibility, the entire region between the two IS1 sequences was cloned as an 18 kb HindIII-BamHI restriction fragment in pUC8 giving plasmid pMO1. A number of derivatives of pMO1, in which aerobactin genes were tagged with a kanamycin resistance gene, were prepared in order to assess the ability of both IS1s to promote the formation of cointegrates with pCJ105, an F derivative devoid of insertion sequences. Mating-out assays indicated that both flanking IS1s were active in cointegrate formation at detectable frequencies. In some cases, the cointegrates could be resolved, the final result being a transposition-like event for the entire aerobactin system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00339620
    Library Location Call Number Volume/Issue/Year Availability
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    Paper (German National Licenses)
    Fulltext
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