Cloning of foot-and-mouth disease virus integrin receptor beta1 subunit and antibody production to its ligand-binding domain.
- Author:
Ping DU
1
;
Youjun SHANG
;
Junwu MA
;
Yanyu HE
;
Xiaolin SUN
;
Xiangtao LIU
Author Information
1. Key Laboratory of Animal Virology, Ministry of Agriculture/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, Lanzhou 730046, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
biosynthesis;
Cattle;
Escherichia coli;
genetics;
metabolism;
Foot-and-Mouth Disease Virus;
physiology;
Integrin alpha1beta1;
biosynthesis;
genetics;
immunology;
Ligands;
Protein Binding;
Protein Interaction Domains and Motifs;
Rabbits;
Receptors, Virus;
genetics;
metabolism;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
immunology
- From:
Chinese Journal of Biotechnology
2008;24(5):874-880
- CountryChina
- Language:Chinese
-
Abstract:
We produced beta1 gene which is about 2400 bp by reverse transcription polymerase chain reaction (RT-PCR) from bovine trachea, reclaimed and purified, then cloned the amplified fragment to pGEM-T easy vector, confirmed by sequencing. The immune-dominant epitope of beta1 gene was chosen by computer analysis and then syncretized ligand-binding domain from 346 bp to 843 bp of ecytoplasm with six histidine, expressed LBD protein massly in E. coli BL21 (DE3), and identified by SDS-PAGE. The fusion protein was purified with Ni-NTA affinity chromatography and immunized New Zealand rabbits preparing of its polyclonal antibody, the specific antibody titer was above 1:12,800 detected by indirect ELISA, the result of Western blot showed that this antibody could be recognized by LBD fusion protein.
