粉尘螨变应原derf2、derf5基因的克隆表达与免疫学初步分析

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1、粉尘螨变应原IX-rt2、D℃m基因的克隆表达及免疫学的初步研究Cloning，ExpressionandPrimaryImmunologicStudyofDerf2，Derf5twoAllergensfromDermatophagoidesFarinaeSubject：PathogenicbiologyMaster：ZhanghaoAdvisor：ZhengXiaoyingDepartmentofParasitology,ZhongshanSchoolofMedicine，SunYat—SenUniversity,Guangzhou，510080AbstractWitht

2、hechangeoflifestyleandpollutionofenvimnn舱nt,theprevalencerateofallergicdiseaseskeepsrising．Allergicdiseasesnotonlydoharmtopeoplehealth,butalsobringgreateconomicburdentofamiliesandsociety．Allergicdiseasesinducedbydustmitesarcthemostcommonallergies．DustmitesarethoughttObemajorcontentsofhouse

3、allergensandthestrongestallergenssincethesixtiesofthe20nIcentury．Mostasthma，allergicrhinitisandatopicdermatitisarerelatedtodustmites．Allergicdiseasesdamagepatientslhealthheavily,however,thereisstillnosatisfieddiagnosticagentsandtherapies．Inourcountry,thecrudeextractfromdustmitesisalwaysado

4、ptedfortreatment,whileitincludesmanykindsofallergicandnonallergiccontentsandisdifficulttoIV粉尘螨变应原Derf2、Derf5肇园的克隆表达及免疫学的初步研究standardme．Sideeffectsmayresultsfromimmunotherapywithcrudemiteextract，suchasallergyandasthma．Furthermore，immunotherapywithcrudemiteextractisdefectivebecausetheallerge

5、nssupplyisdifficult，theallergicproteinsarehardtostandardizeandpurify,etc．Therefore，itisveryimportanttostudyanddevelopnewsaferallergenvaccineswimhigh-effect．ObjeetiveIsolationandpurificationofthegroupIIallergenandgroupVallergenfromthedustmites，DermatophagoidesFarinaeforimmunotherapyarediffi

6、cult．InordertoobtainabundantrecombinantallergenproteinDerf2andDerf5forthefurtherstudyOlldevelopmentofnovelhomemadeanti—allergydrugsandproducepurifiedstandardizedrecombinantallergenproteinforimmunotherapy,theprokaryoticexpressionsystemwasestablishedinattempttoefficientexpressrecombinantalle

7、rgenproteinDerf2andDerf5．MethodsThedustmites,D．farinaefromGuangZhouweresettleddownandreproducedinlargescaleinlaboratory．PairsofprimersweredesignedaccordingtOthepublishedDerf2andDerf5sequences．Theopenreadingframe(ORE)ofDerf2andDerf5geneswereamplifiedbyRT-PCRaft