简介:InordertorevealvariationandrevolutionofNPgenesofhumanavianH_5N_1influenzavirusstrains,theNPgeneofahumanavianH_5N_1influenzavirusstraininGuangdongwassequencedandtheglobalNPgenesofstrainswereretrieved.ThesequenceswereanalyzedbyDNAStar5.0,andtheevolu-tionaryspeedwasstudiedwithreferencetotheepidemiologicaldata.ItwasfoundthatNPgenesof45strainsduring1997-2006werehomologicallyclassifiedintothreegroups:strainsin1997-1998,strainsin2004-2005andstrainsfrom2003to2006.Therewere35substitutionsinNPsinallstrainsaccountingforaratioof7.03%(35/498).Anadditionalglycoproteindomain(NGT_(430-432))wasfoundinNPgenesinthestrainsof2003-2006,themutationofN_(370)SinGD-01-06resultedinoccurrenceofonemoreglyco-proteindomain(NES_(368-370)).Inthesynonymousvariation,K_svaluesinNPwere2.03×10~(-5)-2.55×10~(-5)Nt/dandK_avaluesinNPwere1.58×10(-6)-3.10×10~(-6)Nt/d.Theredidn′texistobviouslyselectivepres-sure.Anadditionalglycoproteindomainineverystrainof2003-2006andonemoreinstrainGD-01-06mightchangetheantigenicityofhumanavianH_6N_1influenzavirus.ThevariationonhumanavianH_5N_1influenzastrainsoccurredfrequentlyinthenaturalworld,whichwouldresultinhighprobabilityofhu-man-humantransmissionalongwiththenaturalevolutionofthevirus.
简介:Theimmuneefficiencyofarecombinantadenovirustype5withtype35fibercontainingHIV-1gaggene(rAd5/F35-mod.gag)wasinvestigatedinBALB/cmice,inwhichtherAd5/F35-mod.gagwasfirstlyidentifiedwithPCR,thentransfectedto293cellsandtheinvitroexpressionlevelofGagproteinwasdeterminedbyWesternblottingandindirectimmuno-fluorescentassay.MicewereimmunizedwithintramuscularinjectionsofrAd5/F35-mod.gag,rAd5-mod.gagorDNAandwereboostedafter3weeks.Totesttheeffectofpre-existinganti-viralimmunityonimmunization,micewerealsoinjectedwithAd5-GFPvectorandthenimmunized4and7weekslaterwithAd5/F35-mod.gagvector.TheP24-specificIgGantibodyinseraofimmunizedmicewasdeterminedbyELISAandthespecificcytotoxicTlymphocyte(CTL)responsewasassayedbyintracellularcytokinestaining.ItwasdemonstratedthattherAd5/F35-mod.gagvectorcouldexpressefficientlytheHIVGagproteinin293cellsinvitroandinducestrongHIV-specificimmuneresponsesinvivo.ThestrongestCTLandserumIgGresponseoccurredwhenmicewereimmunizedtwicewithinjectionofrAd5/F35alone,buttheanti-Ad5antibodyafterprimaryinfectionwithadenoviruscouldinhibitthespecificimmuneresponsesinducedbyrAd5/F35vector.ItisconcludedthatsingleimmunizationwithrecombinantadenovirusrAd5/F35-mod.gagcaninducespecificCTLandserumIgGantibodyresponsesinmice,buttheimmunogenicityofrAd5/F35iscomparablyweakerthanthatofrAd5.