Prokaryotic expression of fowl adenovirus serotype 11 Fiber protein and prepara-tion of its polyclonal antibody
10.16303/j.cnki.1005-4545.2024.07.07
- VernacularTitle:血清11型禽腺病毒Fiber蛋白的原核表达及多克隆抗体的制备
- Author:
Shuaifeng LIU
1
;
Yifan MA
;
Xiangqin WANG
;
Xiaoran GUO
;
Xiaona LIU
;
Baishi LEI
;
Wanzhe YUAN
;
Fuxing CHEN
;
Kuan ZHAO
Author Information
1. 河北农业大学动物医学院,河北 保定 071001
- Keywords:
fowl adenovirus serotype 11(FAdV-11);
Fiber protein;
polyclonal antibody;
cross reac-tions
- From:
Chinese Journal of Veterinary Science
2024;44(7):1387-1393
- CountryChina
- Language:Chinese
-
Abstract:
In order to obtain polyclonal antibodies against the fibrillar(Fiber)protein of fowl ade-novirus serotype 11(FAdV-11)and investigate its cross-reactivity to different serotypes of FAdV Fiber,the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vec-tor pET-32a by homologous recombination technology,then the plasmid was transformed into BL21(DE3)receptor cells,and the purified recombinant protein was used as an immunogen to im-munize rabbits to prepare polyclonal antibody after induced expression,and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a,FAdV-8b,and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2)proteins.In conclusion,in this study,we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a,FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diag-nosis of FAdV-11.
