学科分类
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1 个结果
  • 简介:Tocloneandconstructtherecombinantplasmidcontainingthemajoroutermembraneprotein(MOMP)geneofChlamydiatrachomatis(C.trachomatis)andtoexpressthefusionproteininE.coliBL21,theMOMPgenewasamphfiedbypolymerasechainreaction(PCR)fromgenomeofC.trachomatisserovarD.ThefragmentwasclonedintotheprokaryoticexpressionvectorpET-22b(+)afterdigestionwithBamHⅠandNotⅠandtransformedintoE.coliXL1-Blue.RecombinantswereselectedbyenzymedigestionandsequencingandtherecombinantplasmidwithMOMPgenewasthentransformedintoE.coliBL21withIPTGtoexpressthetargetgene.TheexpressionrecombinantproteinswerepurifiedbyNi-NTAaffinitychromatography,andidentifiedbySDS-PAGEandWesternblot.Itwasfoundthata1.2kbMOMPgenewasisolated.TheDNAsequenceofMOMPwasfoundtobejustthesameasthesequencepublishedbyGenBank.ArecombinantplasmidcontainingMOMPgenewasconstructedtoexpressthefusionproteinsinE.coli.SDS-PAGEanalysisshowedthattherelativemolecularweightoftherecombinantproteinwasabout47kDathatwasconsistentwiththetheoreticalpredictedvalue,andthespecificityoftheexpressedproteinwasconformedbyWesternblot.ItconcludedthattheMOMPgenecouldbeexpressedintheprokaryoticsystem,bywhichitprovidedthefoundationforthefuturestudiesonthebiologicalactivitiesofC.trachomatisandforthedevelopmentofvaccineagainstthispathogen.

  • 标签: 沙眼衣原体 外膜蛋白 原核细胞 基因表达 纯化作用 MOMP