携带幽门螺杆菌中性粒细胞激活蛋白活减毒鼠伤寒沙门菌口服重组D

WorldChinJDigestol2004June;12(6):1317-1320ISSN1009-3079CN14-1260/RPOBox2345Beijing100023,ChinaFax:[phone]Email:[e-mail]•Hpylori•ConstructionofanoralrecombinantDNAvaccinestrainofliveattenuatedSalmonellatyphimuriumcarryingHpylorineutrophilactivatingproteinBoSun,MiaoHe,HuaYang,JingJin,Xiao-HuaMan,Yan-FangGong,Zhen-XingTu,Yi-QiDu,Zhao-ShenLiBoSun,HuaYang,JingJin,Xiao-HuaMan,Yan-FangGong,Zhen-XingTu,Yi-QiDu,Zhao-ShenLi,DepartmentofGastroenterology,ChanghaiHospital,SecondMilitaryMedicalUniversity,Shanghai200433,ChinaMiaoHe,Divisionof,MedicalTechnologyandLaboratory(Brigade26),SecondMilitaryMedicalUniversity,Shanghai200433,ChinaSupportedbytheNaturalScienceFoundationoftheMinistryofHealthofChina,No.30170427Correspondenceto:Yi-QiDu,DepartmentofGastroenterology,ChanghaiHospital,SecondMilitaryMedicalUniversity,174ChanghaiRoad,Shanghai200433,[e-mail]:2003-12-17Accepted:2004-08-16AbstractAIM:ToconstructaliveattenuatedSalmonellatyphimurium(S.typhimurium)straincarryingHpylori-neutrophilactivatingprotein(HP-NAP)geneasanoralrecombinantDNAvaccine.METHODS:Bygeneticengineeringmethod,a435bpnapAgene(encodingHP-NAP)wassubclonedintoaneukaryoticexpressionvectorpIRES.AftersequencingandBLASTanalysis,theidentifiedrecombinantplasmidwasthentrans-formedintoaliveattenuatedS.typhimuriumstrainSL7207.RESULTS:Byusingpolymerasechainreaction(PCR)andrestrictionenzymedigestion,arecombinanteukaryoticexpressionplasmidpIRES-napAcontainingnapAgeneofHpyloriwasconstructed,andtherecombinantplasmidwastransformedsuccessfullyintotheliveattenuatedS.typhimuriumstrainSL7207.MostoftheHpylori-napAse-quencesinrecombinantplasmidpIRES-napAwereho-mologizedwiththatofSS1strainreportedbyGenBank,andthehomologyofnucleotideandproteinwas98%,respectively.CONCLUSION:ArecombinantliveattenuatedS.typhimuriumstraincarryingHP-NAPgeneasDNAvaccineissuccessfullyconstructed.TheresultslaythefoundationfordevelopingmultivalentoralDNAvaccineagainstHpyloriinfection.SunB,HeM,YangH,JinJ,ManXH,GongYF,TuZX,DuYQ,LiZS.ConstructionofanoralrecombinantDNAvaccinestrainofliveattenu-atedSalmonellatyphimuriumcarryingHpylorineutrophilactivatingprotein.ShijieHuarenXiaohuaZazhi2004;12(6):1317-1320ααμΩTplacpUCoriAmprf1(+)orinapA(435bp)pBTVectorT4DNALigaseAmprpIRESVectorMCSAIRESMCSBAmprnapApBT-napAXhoIandMluIcuttingMluIXhoIXhoIMluInapApIRESVectorT4DNALigasenapApIRES-napAIRESMCSBAmpr123456789101112bp15005001001500010005002501001319ααβγAB1BlaserMJ,AthertonJC.Helicobacterpyloripersistence:biologyanddisease.JClinInvest2004;113:321-3332SandersMK,PeuraDA.Helicobacterpylori-AssociatedDiseases.CurrGastroenterolRep2002;4:448-4543ZhangJ,ChenXL,WangKM,GuoXD,ZuoAL,GongJ.Rela-tionshipofgastricHelicobacterpyloriinfectiontoBarrett’sesophagusandgastro-esophagealrefluxdiseaseinChinese.WorldJGastroenterol2004;10:672-6754MaoYF,YanJ.Constructionofprokaryoticexpressionsys-temofureBgenefromaclinicalHelicobacterpyloristrainandidentificationoftherecombinantproteinimmunity.WorldJGastroenterol2004;10:977-9845BaiY,LiLR,WangJD,ChenY,JinJF,ZhangZS,ZhouDY,ZhangYL.ExpressionofHelicobacterpyloriHsp60proteinanditsimmunogenicity.WorldJGastroenterol2003;9:2711-27146QiaoW,HuJL,XiaoB,WuKC,PengDR,AthertonJC,XueH.cagAandvacAgenotypeofHelicobacterpyloriassociatedwithgastricdiseasesinXi’anarea.WorldJGastroenterol2003;9:1762-17667JiangZ,HuangAL,TaoXH,WangPL.ConstructionandcharacterizationofbivalentvaccinecandidateexpressingHspAandM(r)18,000OMPfromHelicobacterpylori.WorldJGastroenterol2003;9:1756-17618LaiYC,WangTH,HuangSH,YangSS,WuCH,ChenTK,LeeCL.DensityofHelicobacterpylorimayaffecttheefficacyoferadicationtherapyandulcerhealinginpatientswithactiveduodenalulcers.WorldJGastroenterol2003;9:1537-15409UemuraN,OkamotoS,YamamotoS.Hpyloriinfectionandthedevelopmentofgastriccancer.KeioJMed2002;51(Suppl2):63-6810LeeSK,LeeYC,ChungJB,ChonCY,MoonYM,KangJK,ParkIS,SuhCO,YangWI.Lowgradegastricmucosaassociatedlymphoidtissuelymphoma:treatmentstrategiesbasedon10yearfollow-up.WorldJGastroenterol2004;10:223-22611Al-AkwaaAM,SiddiquiN,Al-MoflehIA.Primarygastriclymphoma.WorldJGastroenterol2004;10:5-1112WangKX,WangXF,PengJL,CuiYB,WangJ,LiCP.Detec-tionofserumanti-HelicobacterpyloriimmunoglobulinGinpatientswithdifferentdigestivemalignanttumors.WorldJGastroenterol2003;9:2501-250413LanJ,XiongYY,LinYX,WangBC,GongLL,XuHS,GuoGS.Helicobacterpyloriinfectiongeneratedgastriccancerthroughp53-Rbtumor-suppressorsystemmutationandtelomerasereactivation.WorldJGastroenterol2003;9:54-5814WangRT,WangT,ChenK,WangJY,ZhangJP,LinSR,ZhuYM,ZhangWM,CaoYX,ZhuCW,YuH,CongYJ,ZhengS,WuBQ.Helicobacterpyloriinfectionandgastriccancer:evi-dencefromaretrospectivecohortstudyandnestedcase-con-trolstudyinChina.WorldJGastroenterol2002;8:1103-110715GuoDL,DongM,WangL,SunLP,YuanY.Expressionofgastriccancer-associatedMG7antigeningastriccancer,pre-cancerouslesionsandHpylori-associatedgastricdiseases.WorldJGastroenterol2002;8:1009-101316XueFB,XuYY,WanY,PanBR,RenJ,FanDM.AssociationofHpyloriinfectionwi