ULBP4原核表达、生物学功能鉴定及其单克隆抗体制备及初步鉴定

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1、ULBP4原核表达、生物学功能鉴定及其单克隆抗体制备及初步鉴定作者：曹伟，郝志勇；希卩雪艳孔燕马驰崔莲仙何维毕业论文【关键词】;ULBP4；.NKG2D；,IFN毕业论文ExpressionandbiologicalfunctionofULBP4inRosettagamiTMB(DE3)andmAbpreparation毕业论文[Abstract]；AIM：ToexpresshumanULBP4inRosettagamiTMB(DE3)andtopreparemonoclonalantibodyagainstULBP4fortheresea

2、rchofY8Tcellsrecognitionmechanism・METHODS：DNAfragmentsofULBP4werederivedfromH08910RNAbyreversetranscriptasepolymerasechainreaction(RTPCR)・Thefragmentsencodingtheformer225aminoacidsofULBP4wereclonedintoHistagfusionproteinexpressionvectorpET22b(+).TheCHistagfusionULBP4(225a)

3、proteinwasexpressedininclusionbodyULBP4原核表达、生物学功能鉴定及其单克隆抗体制备及初步鉴定作者：曹伟，郝志勇；希卩雪艳孔燕马驰崔莲仙何维毕业论文【关键词】;ULBP4；.NKG2D；,IFN毕业论文ExpressionandbiologicalfunctionofULBP4inRosettagamiTMB(DE3)andmAbpreparation毕业论文[Abstract]；AIM：ToexpresshumanULBP4inRosettagamiTMB(DE3)andtopreparemonoclo

4、nalantibodyagainstULBP4fortheresearchofY8Tcellsrecognitionmechanism・METHODS：DNAfragmentsofULBP4werederivedfromH08910RNAbyreversetranscriptasepolymerasechainreaction(RTPCR)・Thefragmentsencodingtheformer225aminoacidsofULBP4wereclonedintoHistagfusionproteinexpressionvectorpET

5、22b(+).TheCHistagfusionULBP4(225a)proteinwasexpressedininclusionbodyandpurifiedstepbystepaccordingtomanufactory"sprotocolandrenaturedinbagfilter.cellswasevaluatedbysecretionexperiments.ULBP4wasusedasanantibodiesbymeansofIt，sfunctionaleffectonNKNKG2DbindingassayandIFNYProka

6、ryoticexpressedhumanantigentopreparemonoclonaltheBlymphocytehybridomatechnique.RESULTS：ULBP4recombinantproteincanstimula.teNKcellstosecreteIFNy.ThroughPEGfusionandscreeningbylimiteddilution,weobtainedfourstrainsofhybridomacellssecretingantiULBP4antibodies.CONCLUSION：Thefus

7、ionproteinwasexpressedsuccessfullyandfunctional.Atthesametime,theantiULBP4mAbswerepreparedsuccessfully.Bothofthemprovideaplatformforfurtherresearch・毕业论文[Keywords]ULBP4；NKG2D；IFNY；RosettagamiTMB(DE3)E.coli；Expression；Biologicalfunction；monoclonalantibody毕业论文［摘；要］;目的：在大肠杆菌中表

8、达ULBP4重组蛋白，并制备其特异性的单克隆抗体（mAb）,；为V8T细胞对ULBP4的识别机制研究奠定基础。方法：根据GenBank中ULBP4的基因序列设计相应的引物,用RT