B-cell Epitopes Prediction And Identification Of H And F Protein Of Pprv And Its Immunological Characteristics Analyzed

Peste des petits ruminants (PPR) is an acute highly contagious and economically important viral disease of small ruminants,especially goats and sheep,with morbidity and mortality rates as high as 100% and 90%,respectively. Peste des petits ruminants is popular in the world also a serious threat to the sheep industry of China. Therefore, it is very necessary to do some researches on the virology , epidemiology, clinical characters, pathology, diagnose and vaccine development of the disease.B cell epitopes of PPRV H and F antigen were predicted using accessibitity and flexibility schemes,associated with antigenicity,secondary structure and multiple sites prediction,Ten antigen peptides have been designed and synthesized,iELISA detection showed that the ten peptides could react with positive sera of PPRV.The ten peptides were previously conjugated with BSA ,and then ten conjugants were mixed with the same concentration and immunized rabbits.Rabbits sera were collected after the third immunization to evaluate for their specific IgG antibodies against not only their respective homologous peptide but also antibody degree for 75?1 strain and for their capacity to neutralize the virus of Nigeria 75?1.The results showed that these ten synthetic mimotope peptides were antigenic.The sera could neutralize the virus of Nigeria 75/1 in vitro.Ten B cell antigen epitopes from the sequence of H and F protein gene were predicted according to the computer software ,selected by indirect-ELISA ,then synthesized the ten epitopes and an auniversial helper T lymphocyte epitope one by one,these epitopes are comparted by Glycin to keep independen,the recombinant multi-epitope antigen gene,which was inserted into the prokaryotic expression vector PET-30(a),the recombinant PET-30(a) was transformed into E.coli BL21(DE3) and induced for expressing,the molecular weight of the protein was about 19kD;The purified protein could reacted with positive sera against PPRV by indirect-ELISA. The results showed that the recombinant plasmid has been successfully constructed and protein can be highly expressed,the fusion protein could interact with standard positive serm of pprv specially.The recombinant proteins purified were used to immunize mouse, and the specific antibody was assayed by indirect ELISA , the results showed that the fusion protein could interact with the virus of Nigeria 75?1. It may be used as a potential candidate of epitope vaccine of peste des petits ruminants.