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Characterization of the Citrobacter freundii phoE gene and development of C. freundii-specific oligonucleotides
Authors:Gonnie Spierings  Corrine Ockhuijsen  Harmen Hofstra  Jan Tommassen
Affiliation:Department of Molecular Cell Biology, University of Utrecht, Netherlands.
Abstract:The phoE gene of Citrobacter freundii, encoding a pore-forming outer membrane protein, was cloned and its nucleotide sequence was determined. The homologies in terms of identical amino acids between the C. freundii PhoE protein and those of Escherichia coli, E. cloacae and Klebsiella pneumoniae were 90%, 86% and 84%, respectively. Two synthetic oligonucleotides, corresponding to hypervariable, cell surface-exposed regions of the protein, were tested for their specificity in polymerase chain reactions. They were specific for the species C. freundii, i.e., no reaction was detected with 35 non-C. freundii strains tested, including 17 Salmonella, two C. amalonaticus and three C. diversus strains, whereas all five C. freundii strains tested were correctly recognized.
Keywords:Citrobacter freundii    Pore protein    PhoE protein    pho Regulon    DNA probe    Polymerase chain reaction
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