蘿卜過氧化物酶基因RsPrx1在酵母中的表達(dá)、純化及鑒定.pdf

1、河南師范大學(xué)碩士學(xué)位論文蘿卜過氧化物酶基因RsPrx1在酵母中的表達(dá)、純化及鑒定姓名：李芳軍申請(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：王林嵩20090601ABSTRACTPeroxidasewidelyexistsinthebiosphereTheycanbefoundinfungi，bacteria，animalsandplantsThecurrentstudyshowedthatperoxidaseinplantspl

2、aysmainlytwoles，whicharerelativetoplantmorphogenesis，growthanddevelopment；furtheHnoreperoxidasecanhelpplantsagainstcertainbadenvrionment，includingdrought，coldsaltanddiseaseItisoneofthemostimportantprotectionenzymesofplan

3、tInthisstudygsPrxlcDNAwasamplifiedbyreversetranscriptionpolymerasechainreaction(RTPCR)fromthepeelofChineseredradishroot(RaphanussativusVUF刀堙D廠BrassicaceaeL)ThefulllengthofRsPmlcDNAwassequencedandconfnmedThenitwassubclone

4、dint0PichiapastorisexpressionvectorpPIC9KHsuccessfullyRsPrxlgeneWasintegratedintothegenomeofPichiapastor／sWhentherecombinantswereinducedbyBMMYmedium，secretedactivlyPODcouldbetestedStudieshaveshownthatdifferentculturecond

5、itionswouldeffecttheexpressionofforeigngenesinPichiapastorisThisstudyoptimizedthefermentationconditions：BMGYandBMMYmedium，74pH，05％methanol，26。C，aerationaroa15cm2InordertogetsecretedproteinRsPrxlefficientlyweestablishedan

6、ewwayWhenthesecreted睨lzynlereachedthegreatestactivity舭shinducedmediumWaschangedBytestingtheactivityofPODinculturesupematantwhichWasinducedbyBMMYmedia，thetotalactivityofPODinthiswayWas259timesthanthatfTomthegeneralculture

7、method(onlyaddedYP)SDSPAGEshowedthatintheinducementofmethanol，secretedrecombinantRsPrxlhasbeensuccessfullyexpressedinPichiapastoris，andthemassoftherecombinantRsPrxlWasabout37KD，whichisthesameasthevalueofcalculatingtheani

8、moacidsequenceTheresultofNaturePAGEhasprovedthattherecombinantRsPrxlprocessedactivityWhentherecombinantswereculturedinthemediumcontainedNaClorH202，meycouldshowthesaltandoxidationtoleranceInthisstudyanefficientlyextracell

9、ularexpressionsystemofRsPrx1inthehoststrainPichiapastorisyeastGS115wasestablishedwhichunderliesthebaseoflargescaleproductionandfunctionalanalysisKEYWORD：redradish，RsPrxlgene，recombinantexpressionvectorPichiapastoris，indu