Expression and purification of an adhesive protein of rabbit Pasteurella multocida C51-3 and detection of its antigenicity / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1446-1453, 2008.
Artigo
em Chinês
| WPRIM
| ID: wpr-275365
ABSTRACT
The cp36 gene encoding an adhesive protein was amplified by PCR from genomic DNA of rabbit P. multocida C51-3 strain, and cloned into the pMD18-T vector and then sequenced. The mature adhesive protein without a signal peptide of cpm36 gene was amplified by PCR from the recombinant plasmid pMD18-cp36, then cloned into the prokaryotic expression vector pQE30 to provide a recombinant plasmid pQE30-cpm36. The recombinant protein of CPM36 was produced in Escherichia coli M15 harboring the recombinant plasmid pQE30-cpm36 by IPTG induction, and the recombinant protein purified by the affinity chromatography with Ni(2+)-NTA resin. The sequence analyses showed that the ORF of cp36 gene was 1032 bp in length, and DNA homology of the cp36 genes between the C51-3 strain and the previously reported different serotype strains of P. multocida in GenBank was 76.9 to 100%. The SDS-PAGE analyses revealed a single fusion protein band with a molecular weight of 37 kD, and the Western blotting analysis demonstrated that the recombinant protein CPM36 and native 36 kD protein of C51-3 were recognized specifically by an antiserum against the recombinant protein, suggesting that the recombinant protein is an antigenic protein.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes de Fusão
/
Química
/
Pasteurella multocida
/
Clonagem Molecular
/
Adesinas Bacterianas
/
Alergia e Imunologia
/
Escherichia coli
/
Genética
/
Metabolismo
/
Microbiologia
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2008
Tipo de documento:
Artigo
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