Brucella melitensis 16M: Characterisation of the galE gene and mouse immunisation studies with a galE deficient mutant

Liljana Petrovska, R. Glyn Hewinson, Gordon Dougan, Duncan J. Maskell, Martin J. Woodward*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Citations (SciVal)

Abstract

The galE gene of Streptomyces lividans was used to probe a cosmid library harbouring Brucella melitensis 16M DNA and the nucleotide sequence of a 2.5 kb ClaI fragment which hybridised was determined. An open reading frame encoding a predicted polypeptide with significant homology to UDP-galactose-4-epimerases of Brucella arbortus strain 2308 and other bacterial species was identified. DNA sequences flanking the B. melitensis galE gene shared no identity with other gal genes and, as for B. abortus, were located adjacent to a mazG homologue. A plasmid which encoded the B. melitensis galE open reading frame complemented a galE mutation in Salmonella typhimurium LB5010, as shown by the restoration of smooth lipopolysaccharide (LPS) biosynthesis, sensitivity to phage P22 infection and restoration of UDP-galactose-4-epimerase activity. The galE gene on the B. melitensis 16M chromosome was disrupted by insertional inactivation and these mutants lacked UDP-galactose-4-epimerase activity but no discernible differences in LPS structure between parent and the mutants were observed. One B. melitensis 16M galE mutant, Bm92, was assessed for virulence in CD-1 and BALB/c mice and displayed similar kinetics of invasion and persistence in tissues compared with the parent bacterial strain. CD-1 mice immunised with B. melitensis 16M galE were protected against B. melitensis 16M challenge.

Original languageEnglish
Pages (from-to)21-36
Number of pages16
JournalVeterinary Microbiology
Volume65
Issue number1
Early online date29 Jan 1999
DOIs
Publication statusPublished - 23 Feb 1999

Keywords

  • Brucella melitensis
  • galE
  • LPS
  • Mouse immunisation
  • UDP-galactose-4-epimerase
  • Blotting, Southern
  • Brucellosis/immunology
  • Molecular Sequence Data
  • Colony Count, Microbial
  • Bacteriophage P22
  • Mice, Mutant Strains
  • Cloning, Molecular
  • Polymerase Chain Reaction
  • Female
  • DNA, Bacterial/chemistry
  • Amino Acid Sequence
  • Lipopolysaccharides/analysis
  • Brucella melitensis/enzymology
  • Random Allocation
  • Sequence Analysis, DNA
  • Spleen/microbiology
  • Sequence Alignment
  • Animals
  • UDPglucose 4-Epimerase/analysis
  • Mice
  • Mice, Inbred BALB C
  • Mutagenesis, Insertional
  • DNA Primers/chemistry
  • Genes, Bacterial/immunology

Fingerprint

Dive into the research topics of 'Brucella melitensis 16M: Characterisation of the galE gene and mouse immunisation studies with a galE deficient mutant'. Together they form a unique fingerprint.

Cite this