The Exploratory Development Of The Recombinant Of Porcine Protein Of IFN-α Against PRRSV

From the recombinant expression vectors of pGEXIFN-α,which Constructed and appraised correctly, The fusion protein were expressed after the recombinant expression vectors transformed in BL21 of E.coli. There was a strip occurring at the location labled 40.2 kD when expression products were identifited by SDS-PAGE. The strip was specific, because its molecular weight was just the same as the tag in the pET32a of procaryon expression vector and the protein deduced according to porcine IFN-αgene. Optimizeing to the concentration of IPTG and the time of IPTG induceing, the produces of IFN-αincreased. The proteins were purityed by NTA and had only one strip by SDS-PAGE. The expression of the porcine IFN-αgene quantity was twenty percent of the bacterium total protein. Purifing the porcine protein of IFN-α, Using the different density imidazole buffer solution of NTA, the protein of the porcines of IFN-αwere adsorption and purification. By SDS-PAGE, the results indicated that the porcine proteins of IFN-α, the purity was highest,by 400 mM density imidazole buffer solution of NTA adsorbed. The anticipeated aim of the protein purification was achieved. So the purified porteins may use in the anti-virus activity test.By inhibiting the cytopathic effect, the porcine protein of IFN-αwas detected, on the Marc-145 cell, the result indicated that activity of recombinant of porcine protein of IFN-αagainst PRRSV was 5.0×103 U/mg. The experiment of the porcine protein of IFN-αsuppressed the PRRSV in vitro. The experiment obtained that the smallest virulent dentisty to the Marc-145 cell was 0.20mg/ml. The result of the experiment indicated that the porcine protein of IFN-αhad the function of the inhibitoried to anti-PRRSV in vitro. But the functions were not too obvious, by diluting the porcine protein of IFN-αat different concentration gradient, using the inhibiting the cytopathic effect, the function of the porcine protein of IFN-αwere to be dected. The density of the smallest of the blocked denstity was 0.14mg/ml.The results of the recombinant of porcine protein of IFN-αagainst PRRSV in vivo indicated: using the recombinant of porcine protein of IFN-αand the medicines could treat the desease pigs effectively, reducing the treatment course 2?3d, the cure rate was higher than the group of only using the medicine, and the group used the recombinant of porcine protein of IFN-αwas not easy to recur. Through the prevention of the experiment of the recombinant of porcine protein of IFN-α, which studied the diarrhea experimental, the result indicated : after being weaned 14d, comparied to the experimental group and comparisonⅡ, the experimental group was more lower than the comparison group, which was possibley related to the result of the organism immunologic function enhancement; to the antibody titer's and the masculine gender rate of swine plague's immune body was enhanced .