[Cloning of foot-and-mouth disease virus integrin receptor beta1 subunit and antibody production to its ligand-binding domain].
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.
Du
Ping
P
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.
Shang
Youjun
Y
Ma
Junwu
J
He
Yanyu
Y
Sun
Xiaolin
X
Liu
Xiangtao
X
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002417:Cattle; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029544:Foot-and-Mouth Disease Virus / Q000502:physiology*; D039222:Integrin alpha1beta1 / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology; D008024:Ligands; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011817:Rabbits; D011991:Receptors, Virus / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D039222:Integrin alpha1beta1; D008024:Ligands; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins