Screening And Identification Of Affinity Peptide To S_Δ16 Protein Of Procine Epidemic Diarrhea Virus

The test successfully constructed a recombinant expression vector pcDNA3.1-S_Δ16. Therecombinant pcDNA3.1-S_Δ16plasmid was transfected into cells mediated. The positive cell clones werescreened with G418. The results showed that the structure of recombinant pcDNA3.1-S_Δ16vector wascorrect vertified by sequencing. The established cell line was able to express S_Δ16mRNA and proteinidentified by RT-PCR and indirect immunofluorescence, respectively.Then in order to generate polyclonal antibodies of the M13Phage, one M13of Ph.D.-12TMPhageDisplay Peptide Library was amplified and used to immunize a rabbit to generate polyclonal antibody.Indirect phage-ELISA analysis showed that the anti-M13antibody had a high titer and the titer of thepolyclonal antibody was approximately1:1048576. Compared this polyclonal antibody with a Rabbitpolyclonal antibody （Rb pAb） to M13Bacteriophage Coat Proteins （commercialization）, the bindingactivity of the produced polyclonal antibody to the identical target was as good as that of the Rb pAb toM13Bacteriophage Coat Proteins. In the study, polyclonal antibody to this M13of phage library wassuccessfully generated and such phage polyclonal antibody is important material for functional analysiswith phage.The stable expression cell lines were screened three rounds through the Ph.D.-12TMPhage displaylibrary. Pick out random phage monoclones from peptide libraries screened, and analyze thenucleotidesequence. The results showed that the sequence-AETVESCLAKSH-was both found in the surface andinternalization. The peptide have the highest repeatability to the target cell was synthesized. Theinhibition of the phage bearing the peptide to PEDV was evaluated in Vero-E6cell byImmunofluorescence and Real time quantitative PCR. The results indicated PEDV and peptideincubation led to a decreased infectivity of PEDV.