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1、___________________________________________________________________________http://www.paper.edu.cn日本血吸虫DNA多价疫苗pVIVO2SjFABP－23的构建及其保护性免疫11※11222李春艳余龙江刘智朱路朱晓华胡媛石佑恩摘要：目的：构建日本血吸虫多价DNA疫苗pVIVO2SjFABP－23，并观察其在小鼠抗血吸虫感染中的免疫保护作用。方法：根据SjFABP和Sj23的基因序列合成两对特异性引物,利用重组
2、PCR技术将日本血吸虫SjFABP和Sj23的基因片段进行拼接,得到融合基因SjFABP-23,再将该片段重组于pGEM-T载体中,进行PCR扩增及DNA序列测定。然后经双酶切将目的片段SjFABP-23克隆到pVIVO2的一个多克隆位点上转化E.coliGT110获得重组质粒pVIVO2SjFABP－23，免疫小鼠进行免疫保护性实验。结果：PCR扩增出一条大小约为1.1Kb的目的片段。免疫小鼠获得52.14%减虫率(P<0.05)和60.93%的减卵率（P<0.01）。结论：PCR成功扩增SjFABP
3、－23的基因片段，血吸虫DNA多价疫苗pVIVO2SjFABP－23构建成功，该疫苗在小鼠抗血吸虫感染中具有一定的免疫保护作用。关键词：日本血吸虫；23KDa膜蛋白；脂肪酸结合蛋白；多价DNA疫苗；免疫保护CONSTRUCTIONOFTHEMULTIVALENTDNAVACCINEpVIVO2SjFABP－23OFSCHISTOSOMAJAPONICUMANDSTUDIESONITSIMMUNOPROTECTIONINMICE11※1LIChun-YanYULong-JiangLIUZhietal(Co
4、llegeofLifeScience&Technology,HuazhongUniversityofScience&Technology,Wuhan430074,China；TongjiMedicalCollege,HuazhongUniversityofScienceandTechnology,Wuhan430030,China)Abstract:ObjectiveToconstructandpreparethemultivalentDNAvaccinepVIVO2SjFABP－23andobserv
5、eitsimmunoprotectionagainstSchistosomajaponicuminmice.MethodsTwocouplesofspecificprimersweredesignedaccordingtogenesequencesofSjFABP(fattyacidbindingprotein)andSj23（23KDamembraneprotein）,twofragmentswereobtainedandsplicedthroughalinkersequencebyrecombina
6、tionPCR.FusionfragmentformedwasinsertedintopGEM-TEasyvector,andidentifiedbyrestrictionanalysisandsequence,digestedwithBamHIandEcoRI,clonedintoexpressedplasmidpVIVO2/MCSandtransformedinE.coliGT110,recombinationplasmidpVIVO2SjFABP－23wasextracted,purifiedan
7、dinoculatedintoBALB/cmicebyintramuscularinjection.MicewerechallengedbySchistosomajaponicumcercariaeandthenkilled.Adultwormandeggwerecounted,respectively.ResultsElectrophoresisofSjFABP-23on0.7%agarosegelshowedanobviousbandof1.1Kb,whileonesofSjFABPandSj23w
8、ere440bpand670bp,respectively.RecombinationplasmidpVIVO2SjFABP－23wasthenconstructed。Protectiveexperimentsappearedthatwormreductionratewas52.14%(P<0.05),andeggsreductionratewas60.93%(P<0.01)whencomparedwiththecontrolgroupin
