一種編碼擬南芥鈣調(diào)素結(jié)合蛋白基因的載體構(gòu)建及其初步表達(dá).pdf

1、華中師范大學(xué)碩士學(xué)位論文一種編碼擬南芥鈣調(diào)素結(jié)合蛋白基因的載體構(gòu)建及其初步表達(dá)姓名：肖逸群申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：植物細(xì)胞工程指導(dǎo)教師：楊萬(wàn)年20070612⑥碩士學(xué)位論文MASTER’STHESISAbstractCamodulin(CaM)‘a(chǎn)sallimportantsecondmessengerThecytosolicfreecalciumlevelleadstobindingofCa2sensorproteinanditsta

2、rgetproteinsinitiatingavarietyofeellttlarprocessesCaMisamajorcalciumsensorproteinexistinalleukaryoticcells，includingin#antsInplantceils，therewerethreeinlracellularsignaltransducfionpathways：Gproteinandphospholipidsignali

3、ngsystems，cyclicnucleosidecAMPandcGMPsignalingsystemsandC擴(kuò)signalingsystemCaMplaysimportantrolesinCa2signalingpathwaydifferentCaMisfformsinteracted晰也itstargetproteinsInArabidopsis，CaMl一CaM7weretypicalCaMisoformproteinsUse

4、pETprokaryoticexpressionsystem，inductionandexpressionthevectorpETCaM2byIPTGinEcoliBL21(DE3)SeparationandbiotinylatedCaMebychromatographyforthepreserveinordertoanalysisoftheinteractionofourtargetproteinandCaMAccordingtoth

5、ebioinformaticsanalysisinArabidopsis，Wefoundagene(At5962390)encodingforcalmodulinbindingproteinFurtheranalysisshowedthatitcontainedtwomotifsforC一dependentbinding：1—510and1814onemotifIQforC礦independentbindingAt5962390，asa

6、template，startatCterminal，amplifieddifferentcDNAscontaindifferentdomainsbyPCRConstructiondifferentvectorsbysubcloneTheresultsshowedthatthecDNAswererespectivelyrightamplifiedbyAgarosegelelectrophoresisIdentificationofreco

7、mbinantplasmidsshowedcorrectbydigestedwi血enzymeandbysequencingDuringtheprocessofstudyingtheconditionsofproteinexpressionwefoundnoexpressionatdifferentIPTG—inducedconcentrationanddifferenttemperatureonthedifferentfragment

8、sInthesameexperimentalconditionswe峨thegenesmall4S0bpfiagmentofrecombinantexpressionvectorsuccessfullyonexpressionThisworklaidthefoundationforourfilrtherstudyKeywords：Arabidopsisthaliana；bioinforrnaticsanalysis；calciumbin