Cloning and expression of duck C4BPα and verification of its interaction with Riemerella anatipestifer / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 693-699, 2020.
Artículo
en Chino
| WPRIM
| ID: wpr-826907
ABSTRACT
To study the interaction between C4b-binding protein (C4BP) and Riemerella anatipestifer (RA), we cloned duck C4BPα, conducted prokaryotic expression and prepared the polyclonal antibody by immunizing mice. Then indirect immunofluorescence assay and dot blotting hybridization assay were used to verify the interaction between C4BP and RA. The full length of duck C4BPα nucleotide sequence was 1 230 bp, with the highest similarity to chicken C4BPα (82.1%). Phylogenetic tree analysis showed that duck C4BPα and chicken C4BPα were on the same phylogenetic tree branch and the genetic evolution relationship between them was the closest. C4BPα was efficiently expressed in Escherichia coli BL21 (DE3). The recombinant proteins existed in intracellular soluble form. The titer of polyclonal antibody was more than 110 000 and polyclonal antibodies could specifically recognize the recombinant proteins. The results of indirect immunofluorescence assay and dot blot hybridization assay showed that RA could interact with duck C4BP. The results provide a basis to further reveal the pathogenesis of RA.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Filogenia
/
Química
/
Regulación de la Expresión Génica
/
Clasificación
/
Clonación Molecular
/
Patos
/
Proteína de Unión al Complemento C4b
/
Riemerella
/
Genética
/
Metabolismo
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2020
Tipo del documento:
Artículo
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