Study On Coadministration Of Recombinant Porcine Interferon-alpha And Astragalus Polysaccharide

This test is mainly based on the combination of recombinant porcinealpha interferon and herbal extracts astragalus polysaccharide starting point, throughthe study of the pig alpha interferon gene cloning and expression, purification andcharacterization of recombinant porcine alpha interferon, Interferon and dosage ofastragalus polysaccharide in three aspects. Study the joint between the the pig alphainterferon and Astragalus polysaccharide interaction in anti-viral activity, providetheoretical guidance for the health farm.First, The prokaryotic expression of porcine alpha interferonIn order to study the porcine alpha interferon antiviral agents, the laboratorysuccessfully constructed prokaryotic expression vector the pmd-18T-PoIFN-alphaexpression in E.coli BL21(DE3) porcine alpha interferon, this expression product is afusion protein. SDS-page electrophoresis appears in26.5kd at a strip, it is consistentwith the expected results. Its production was increased after IPTG induction in theexpression of the porcine alpha interferon, the expression level of28%of the totalbacterial protein, it purified porcine alpha interferon degree of purification can bemore than95%in the testing of the antiviral activity.Second, the determination of porcine alpha interferon activity.The determination of activity of the recombinant porcine alpha interferon withthe cytopathic effect inhibition assay on Marc-145cells. The results showed that: themeasured of PRRSV recombinant porcine alpha interferon anti-viral activity of5.0x105u/mg. The recombinant porcine alpha interferon at the cellular level in vitroagainst PRRSV play an effective inhibition of the reference for further used in animaltests.Third, the amount of interferon and Astragalus polysaccharides.Marc-145cells cultured in vitro, the maximum non-toxic concentrations ofastragalus polysaccharides on Marc-145cells measured by the CPE and MTT methodto calculate the half inhibitory concentration.Comprehensive evaluation of astragaluspolysaccharide anti-of PRRSV effect and value, it found the APS and recombinantporcine alpha interferon against PRRSV has obvious synergies.