Cloning and expression of duck C4BPα and verification of its interaction with Riemerella anatipestifer.
- Author:
Delong LI
1
;
Lijuan TAN
1
;
Jiulong GU
1
;
Siyuan WANG
1
;
Ting LIU
1
;
Sihuai CHEN
1
;
Jiye GAO
1
;
Fashu TANG
1
;
Jixiang LI
1
Author Information
- Publication Type:Journal Article
- Keywords: Riemerella anatipestifer; cloning; duck C4 binding protein α; prokaryotic expression
- MeSH: Animals; Cloning, Molecular; Complement C4b-Binding Protein; chemistry; genetics; metabolism; Ducks; classification; genetics; microbiology; Gene Expression Regulation; Mice; Phylogeny; Riemerella; metabolism
- From: Chinese Journal of Biotechnology 2020;36(4):693-699
- CountryChina
- Language:Chinese
- 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 1:10 000 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.
