Molecular heterogeneity of plpE gene in Indian isolates of Pasteurella multocida and expression of recombinant PlpE in vaccine strain of P. multocida serotype B: 2.
10.4142/jvs.2010.11.3.227
- Author:
Ajay Pratap SINGH
1
;
Satparkash SINGH
;
Rajeev RANJAN
;
Santosh Kumar GUPTA
;
Vijendra Pal SINGH
;
Bhaskar SHARMA
Author Information
1. Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Bareilly- 243122, UP, India.
- Publication Type:Original Article
- Keywords:
Haemorhhagic septicaemia;
Pasteurella multocida;
PlpE
- MeSH:
Animals;
Bacterial Outer Membrane Proteins/*genetics/immunology/metabolism;
Base Sequence;
Blotting, Western;
Cattle;
Cattle Diseases/*microbiology;
Cloning, Molecular;
Electrophoresis, Polyacrylamide Gel;
Escherichia coli;
*Genetic Variation;
Hemorrhagic Septicemia/microbiology/*veterinary;
India;
Lipoproteins/*genetics/immunology/metabolism;
Molecular Sequence Data;
Open Reading Frames/genetics;
Pasteurella multocida/*genetics/immunology;
Sequence Analysis, DNA;
Sequence Homology;
Serotyping;
Species Specificity
- From:Journal of Veterinary Science
2010;11(3):227-233
- CountryRepublic of Korea
- Language:English
-
Abstract:
Outer membrane proteins of Pasteurella (P.) multocida have been known to be protective immunogens. Pasteurella lipoprotein E (PlpE) has been reported to be an important cross reactive outer membrane protein in P. multocida. The gene encoding the PlpE of P. multocida serotypes A: 3, B: 2 and D: 1 was amplified from the genomic DNA. The amplified products were cloned and the nucleotide sequence was determined. Sequence analysis of the recombinant clones revealed a single open reading frame of 1,011 bp, 1,008 bp and 1,017 bp encoding a protein with a calculated molecular mass of 37.829 kDa, 37.389 kDa and 37.965 kDa for serotypes A: 3, B: 2 and D: 1 respectively. The comparison of the plpE sequence in different capsular types revealed a high degree (>90%) of homology. Furthermore, the plpE gene of Haemorhhagic septicaemia causing serotype (B: 2) was expressed in E. coli and recombinant PlpE was strongly immunostained by antiserum against whole cell antigen, indicating that the protein is expressed in vivo.