30702.植物cna類似物的探尋與ppkl家族新成員bsl4的初步鑒定

1、北京師范大學(xué)碩士學(xué)位論文植物CNA類似物的探尋與PPKL家族新成員BSL4的初步鑒定姓名：宋宏濤申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：生物化學(xué)與分子生物學(xué)指導(dǎo)教師：魏群20090601北京師范人學(xué)碩上學(xué)位論文ThedetectionofCNAanalogueinplantandTheprelimiaryidentification幻ffnwelmemlerofPPKL：BSL4idlicatmnanovelmem幻肚：AbstractThisthes

2、iscontainstwomainsectionincludingninechaptersThefirstsectionisaboutthebiochemicalidentificationandphosphataseactivitydetectionofCNAanalogueinplantThesecondsectionisaboutthepreliminaryidentificationofBSIA，anovelmemberofth

3、ePPKLfamilyinArabidopsisthalianaDetailsaredescribedbelowInthefirstpart，WecarriedouttoexploreCNAanaloguesinplantUsingtwocommonCNsubstrates，pNPPandRIIpeptide，WedetecteditsphosphataseactivityTheresultsshowedthattobacoCaMaff

4、inityproteinrepresentedthedistinctcharacterofCNAphosphataseactivityItsphosphataseactivitycouldbeactivatedbyCa2十／CaM，whilewiththesubstrateofisotopelabelingRIIpeptidesubstrate，itsphosphataseactivitycouldberetrainedbyCsA／Cy

5、PorFK506／FKBPwhichwereconsideredastheCNspecificimmunosuppressiveagentsBytheapproachofTwo—dimensionalgelelectrophoresiscoupledwithwesternblot，weidentifiedoneproteinspotfromtobaccoCaMaffinityproteinsamples，whoseMWandP1were

6、similartoCNA，andweinitiallydeterminedthatitwasapotentialCNAanalogueinplantAscontinueofthefirstpart，thesecondWasdesignedtodetecttheCNAanaloguesinplantThestorystartedwithonenovelgenewhichwasoccasionallyclonedbyUSWecalledit

7、BSL4，whichwasidentifiedasonenovelmemberofPPKLfamilyinArabidopsisthalianaThepreliminaryidentificationreferredtoseveralaspects，suchassequence，molecularweight，isoelectricpoint，andsubcelleularpredict，secondaryandthreedimensi

8、onalstructurepredictWiththecommonsubstrateofproteinphosphatasepNPPinphosphataseactivitytest，weidentifiedthatBSIA’SactivitycouldbesignficantlyactivitedbyMn2十，withnearlyhundredfoldactivationmultiple100uMMn2十couldactivateBS

9、L4’SactivityuptotheapexTheoptimumPHandsaltionintensitywerePH74and100mMsaltconcentrationComparedtoBSL4，BSL4一C’sactivitycouldnotberegulatedbyMn2，Ca2，Ca2CaM，EGTAanditsactivityWasfarfromlowerthanBSIA’SAnyotherbivalentmetalio

10、nsexceptforMnzcouldnotregulateBSL4’SphosphataseactivityInadditionweproducedmonospecificPolyclonalantibodies(PcAb)againstBSL4Byyeasttwo—hybridproteininteractionscreeningweobtainedfourspecificproteins：RPP3B，Serineproteasei