Abstract
Mutagenesis with the transposon Tn916 was used as a strategy to identify genes required for synthesis of the Gal alpha (1-4) beta Gal component of Haemophilus influenzae strain RM7004 lipopolysaccharide. Insertion of Tn916 into an open reading frame (ORF) encoding a protein with 75% homology to the Escherichia coli methionine related protein (Mrp) is described. Mutations in mrp resulted in loss of reactivity with monoclonal antibody (mAb) 4C4, which recognises Gal alpha (1-4) beta Gal, and expression of LPS with a different electrophoretic profile to that of wild-type RM7004. An unexpected feature of this mutation was that it appeared to influence the number of copies of 5'-CAAT-3' present in lic2A, a gene which is also required for biosynthesis and phase variable expression of the Gal alpha (1-4) beta Gal LPS epitope.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibodies, Monoclonal
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Bacterial Proteins / genetics
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Bacterial Proteins / immunology
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Bacterial Proteins / metabolism
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Bacteriological Techniques
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Base Sequence
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Blotting, Southern
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Chromosomes, Bacterial / genetics
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DNA Transposable Elements / genetics*
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DNA, Bacterial / genetics
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Enhancer Elements, Genetic / genetics
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Genes, Bacterial / genetics*
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Genetic Testing
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Haemophilus influenzae / cytology
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Haemophilus influenzae / genetics*
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Haemophilus influenzae / metabolism
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Lipopolysaccharides / biosynthesis*
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Molecular Sequence Data
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Mutagenesis / physiology
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Plasmids
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Polymerase Chain Reaction
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Restriction Mapping
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
Substances
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Antibodies, Monoclonal
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Bacterial Proteins
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DNA Transposable Elements
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DNA, Bacterial
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Lipopolysaccharides