單鏈抗體637與人血清白蛋白融合蛋白的真核表達及功能鑒定.pdf

1、延邊大學碩士學位論文單鏈抗體637與人血清白蛋白融合蛋白的真核表達及功能鑒定姓名：李芳芳申請學位級別：碩士專業(yè)：病原生物學指導教師：孟繁平20070418遼邊尺乍他l憊乍物學0址幀’譬衍論殳^hslnwlEukaI了oticexpressionandfhnctionaI￡haracterizationoffusionproteinofsingIechainVariabIefragment637andhumanscrumaIbuminAb

2、stractobjective：ToexpresstheconslructedrecombjnantgeneofsingIechainvariablefragment(ScFV)637ofanljbodyagainstthemainimmunogenicregion(MIR)ofacetyIchoIinereceptor(AChR)inmyastheniagraVis(MG)andhumanserumaIbuminARerthemole

3、cuIarweightoffusionproteinwasconnmed，theafnnityoffusionpmteintoAChRinhumanmfercos扭lsmusclesurasdetectedMethods：A療erresuscicatingtheJP『c廳脅，搿sm，‘捃GS1l5strainswhichbearsrecombinantgeneepressionvectorthestrongresistanlG418of

4、mulljpIecopiestransfomlantswereseIectedAsinglecolonywasincubatedjnYPDBMGYandBMMYcuIturemediatoinduceexpressioninturnWithininducingexpression3days，lOO％methan01waSaddedtoBMMYevery24hourslOmakethefinaIconcentrationOf1％intot

5、aIculturemediumAftercentrifugingIhesupernacantofexpressionmediumat14000gatroomtemperaturethefusionpro【eininsupernatantwasdetectedprimarilybydO‘blotThemolecuJarweiglltoffusionproteinwasanal，’zedbyCoomassiestainedsodiumdod

6、ecylsult’atepolyacryIamidegeIelectrophoresis(SDSPAGE)TheafnnityoffusionproteintoAChRwasdetectedusingindrectimmunonuorcscencetechnologyResuIts：ThefLlsionproteinwassecrete4inthesupernatanIofexpressionculturemediumItsmolecL

7、IIarwei址11I、、asahoLII974KDTherewas門uorescencebetweenthehumanintercostalsnluscIeceIIsConcIusion：Thefusionpfoeein、vasexpressedfiom尸，f^脅，(“，D，’時GSlI5successfLlllyanditcouldbindtoIheAChRinhumanintercostalsmLIscIesKeywords：si