Cloning, expression and immunity of pilA gene and ompC gene from avian pathogenic Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1561-1567, 2008.
Artículo
en Chino
| WPRIM
| ID: wpr-275321
ABSTRACT
In order to amplify pilA gene and ompC gene of avian pathogenic Escherichia coli (APEC) strain, two pairs of primers were designed according to the GenBank sequences, and a 549 bp pilA gene and a 1104 bp ompC gene were obtained by PCR separately. Sequence analysis indicated that the homology of the nucleotide sequence of AEPC strain to those other reference strains was 98.18% of the pilA gene and 97.28% of the ompC gene. Two expression plasmids pETpilA and pETompC were constructed by inserting pilA gene and ompC gene into the prokaryotic expression vector pET-28a. The two plasmids were transformated into E. coli BL21 separately and two recombinant strains BL21 (pETpilA) and BL21 (pETompC) were obtained. The type 1 fimbraie and the out membrane protein were highly expressed when the recombinant strain BL21 (pETpilA) and BL21 (pETompC) were induced by IPTG Two specific proteins were detected by SDS-PAGE and immunogenicity of the expressed protein was confirmed by Western blotting and ELISA. The expressed fimbraie and OmpC were transformed into vaccine. The protective immune response was proved after the mice were immunized with the two vaccines. The results showed that the recombinant strain BL21 (pETpilA) and BL21 (pETompC) could be as candidate vaccine to provide protective immune response against AEPC infection.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes de Fusión
/
Regulación Bacteriana de la Expresión Génica
/
Clonación Molecular
/
Porinas
/
Vacunas contra Escherichia coli
/
Proteínas de Escherichia coli
/
Proteínas Fimbrias
/
Alergia e Inmunología
/
Escherichia coli
/
Genes Bacterianos
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2008
Tipo del documento:
Artículo
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