Outer Membrane Proteins of
Brucella abortus Vaccinal and Field Strains and their Immune
Response in Buffaloes
Rukhshanda Munir*, M. Afzal1,
M. Hussain2, S. M. S. Naqvi3 and
A. Khanum3
Animal Health Program, Animal Sciences Institute, National
Agricultural Research Centre, Park Road, Islamabad; 1Animal
Sciences Division, Pakistan Agricultural Research Council,
Islamabad;
2Faculty
of Veterinary Sciences,
Pir Mehr Ali Shah Arid
Agriculture University, Rawalpindi;
3Department
of Biochemistry, Pir Mehr Ali Shah Arid Agriculture University,
Rawalpindi, Pakistan
*Corresponding author:
rakhshindamunir@yahoo.com
Abstract
Outer membrane proteins (OMPs) of three strains of
B. abortus i.e. S19, RB51 and a local
field isolate of biotype 1 were isolated through disrupting cells to generate
membranes by centrifugation and sodium lauryl sarcosinate solubilisation of
inner membrane proteins. Distinct OMP profiles of each strain were seen on
SDS-PAGE. SDS-PAGE analysis of S19 and field isolate revealed eight protein
bands in each strain.The OMPs of S19
had molecular masses
89.0, 73.0, 53.7, 49.0, 38.0, 27.0, 22.3, and 17.7 kDa, while
field isolate had OMPs of 151.3, 89.0, 75.8, 67.6, 37.0, 27.0, 24.0 and
19.0 kDa. B. abortus
RB51 yielded 11 OMP bands ranging from 12.5 to 107.1 kDa, with 34.2, 15.8 and
12.5 kDa as additional OMPs.
Western immunoblot analysis using antisera raised against all three strains in
buffaloes indicated an almost similar pattern of immuno-reactive OMPs in S19 and
field strain. Two OMPs of molecular weight 37-38 and 19 kDa were immuno-reactive
in all strains in buffaloes. There is possibility of use of these OMPs in a
recombinant vaccine for B. abortus. A
distinct protein of molecular weight of 151.3 kDa was identified in field strain
but not in both vaccine strains of B.
abortus. Use of this OMP in a diagnostic assay may differentiate between
vaccinated and infected animals.
Key words: Protein
profiling,
Western blot analysis,
Outer membrane proteins, Brucella
abortus,
RB51,
S19,
Buffaloes