Identification of a new iron regulated locus of Salmonella typhi

Andreas J. Bäumler, Renée M. Tsolis, Adrianus W.M. Van Der Velden, Igor Stojiljkovic, Suzana Anic, Fred Heffron

    Research output: Contribution to journalArticle

    129 Scopus citations

    Abstract

    In order to identify genes belonging to the Fur regulon of Salmonella typhi which are absent from Escherichia coli K-12, a plasmid gene bank consisting of 4000 independent clones was screened for Fur regulated promoters using the Fur titration assay (FURTA). DNA probes generated from FURTA positive plasmids were then used for hybridization with chromosomal DNA from S. typhi, Salmonella typhimurium and E. coli. Using these techniques we identified an iron regulated locus present in S. typhi and S. typhimurium but not in E. coli. Further cloning and nucleotide sequence analysis identified two open reading frames, termed iroBC, organized in a typical operon structure. The genes iroBC were located at 4 and 57 centisomes on the physical maps of Salmonella typhi and S. typhimurium, respectively. This region of the S. typhimurium chromosome contains a large DNA loop which is absent from the corresponding area of the E. coli chromosome. Finally, we developed a new method for generation of single copy transcriptional fusions. A suicide vector was constructed, which allows for the generation of chromosomal fusions to the promoterless E. coli lacZYA genes. By integration of this construct at the iro locus we could establish iron responsive expression of iroBC.

    Original languageEnglish (US)
    Pages (from-to)207-213
    Number of pages7
    JournalGene
    Volume183
    Issue number1-2
    DOIs
    StatePublished - Dec 1 1996

    Keywords

    • Fur regulon
    • Pulsed field electrophoretic mapping
    • Salmonella loops
    • Salmonella typhimurium
    • Suicide vector
    • Transcriptional fusion
    • iro
    • β-galactosidase

    ASJC Scopus subject areas

    • Genetics

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  • Cite this

    Bäumler, A. J., Tsolis, R. M., Van Der Velden, A. W. M., Stojiljkovic, I., Anic, S., & Heffron, F. (1996). Identification of a new iron regulated locus of Salmonella typhi. Gene, 183(1-2), 207-213. https://doi.org/10.1016/S0378-1119(96)00560-4