1.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.
Ajay Pratap SINGH ; Satparkash SINGH ; Rajeev RANJAN ; Santosh Kumar GUPTA ; Vijendra Pal SINGH ; Bhaskar SHARMA
Journal of Veterinary Science 2010;11(3):227-233
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.
Animals
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Bacterial Outer Membrane Proteins/*genetics/immunology/metabolism
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Base Sequence
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Blotting, Western
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Cattle
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Cattle Diseases/*microbiology
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli
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*Genetic Variation
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Hemorrhagic Septicemia/microbiology/*veterinary
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India
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Lipoproteins/*genetics/immunology/metabolism
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Molecular Sequence Data
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Open Reading Frames/genetics
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Pasteurella multocida/*genetics/immunology
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Sequence Analysis, DNA
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Sequence Homology
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Serotyping
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Species Specificity