28883.不同蛋白轉(zhuǎn)導(dǎo)域的轉(zhuǎn)導(dǎo)功能初步研究

1、華東理工大學(xué)碩士學(xué)位論文不同蛋白轉(zhuǎn)導(dǎo)域的轉(zhuǎn)導(dǎo)功能初步研究姓名：肖偉申請學(xué)位級別：碩士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：趙健20091230ThePreliminaryStudyonTransductionofDifferentProteinTransductionDomainsAbstractProteintransductiondomain(PTD)，ashortpeptidecarryingotherbiomacromolecul

2、esintocell，isapotentiallynovelandeffectivealderfordiseasetherapyandstudywiththehelpofEGFPandApoptin，weconstructedfiverecombinantplasmids，andthenthefivefusionproteinswereexpressedandpurifiedthepmliminarystudytransductionW

3、asdoneonthethreefusionproteins、析n1EGFPandthepreliminarypharmacologicalactivitystudyWasdoneonthetwofusionproteinswitllApoptintoshowthetransductionofthreeproteintransductiondomainsWefirstsuccessfullyconstructedthreerecombi

4、nantexpressionvectorsseparatelyincludingenhancedgreenfluorescentprotein(EGFP)andspecialproteintransductiondomain(PTD)：EGFPTATpET28a，EGFPhCECSOD—pET28aandEGFPAr97pET28aandthesethreefusionproteinswerehighlyexpressed、析也shak

5、ingflasksandwellpurifiedwitllNiNTAaffinitychromatographypurification：EGFPTATEGFP—hCEC—SODandEGFPAr97wealsosetuplargescaleisolationandpurificationprotocolsforthesethreefusionproteins，thepurityofwhichismorethan90％Thepilots

6、tudyontransductionofthreePTDs、析mEGFPWasdonethestudyconfirmedtheexsiting行ansducfionofthethreePTDs，andindicatedthatthethreePTDSshowedrelativelygoOdtransductiOnin13hrsSecondlytwofusionproteinsseparatelyincludingTATandhC—ECS

7、ODwereexpressed、^ritllshakingflasksandwellpurifiedwitllNiNTAaffinitychromatographypurification：TATApoptinandhC—ECSODApoptinthepurityofthesetwoismorethan90％nepreliminarypharmacologicalactivitystudyshowedthatthetwofusionpr

8、oteinswithApoptinwerebotllprovedhi曲apoptosisactivityagainsttumorcellusinginvitroHeLacellinhibitiontestthisconfirmedtheexsitingtransductionofthetwoPTDs：TATandhC—EC—SODthedifferenceofIC50ofTATApoptinandhCECSODApoptinonHeLc