擬南芥psrp-3基因反義突變體構(gòu)建及其初步生理生化研究.pdf

1、蘭州大學(xué)碩士學(xué)位論文擬南芥psrp3基因反義突變體構(gòu)建及其初步生理生化研究姓名：王耘申請學(xué)位級(jí)別：碩士專業(yè)：生物學(xué)植物學(xué)指導(dǎo)教師：張立新20060501蘭州大學(xué)碩士學(xué)位論文AbstractSixribosomalproteinsarespecifictohigherplantchloroplastribosomesTheseproteinsareplastid—specificribosomalproteins(PSRPs)PSRP一3

2、，onthesmallsubunitandunassociated淅tllrRNAisanevolutionarilyconservedchloroplastribosomalproteinItisthehigherplantorthologueofproteinYCF65Cvcf6sgeneproduct)incyunobacteriaIt、is●proposedthatPSRP一3actsasregulatortoparticipa

3、teinlight—dependentchloroplastproteinsynthesis，howeverthemechanismisnotresolvedyetInourresearchweuseantisensetechnologytostudythephysiologicalandbiochemicalcharacerisicstounderstandthefunctionandmechanismofproteinPSRP3nl

4、emainresultswerereportedasfollowing：1Psrp一3anti—senseeDNAwasamplifiedfromArabidops括thalianabyRTPCRusingapairsofspecificprimerscontaining5’endoftwopoly—linkersXbaIandSadDNAsequenceanalysisindicatesPsrp一3(GeneBankAccession

5、NoATlG68590)consistsof743bp，encodingapolypeptideoft66aminoacidsThefirst44aminoacidsaresignalpeptidesfortargetingtothechloroplast2TheproductofPCRdigestedbyXbaI／SaclwasligatedtopBll21vectorthentherecombinantplasmidpBll21an

6、ti—sensePsrp一3wasobtained。TherecombinantplasmidpBll21一anti—sensePsrp3WasconfirmedbyPCRandsequencealignment3，TherecombinantplantexpressionvectorcontainingpBl121一anti—sensePsrp3wasconstructedandtransferredintoA，tumefaciens

7、EHAl05bytriparentmatingmethodArabidopsisthalianawastranformedwitlltherecombinantAtumefaciensandthetransformantswereidentifiedbyKanresistanceandexaminedbyPCR4Thephysiologicalandbiochemicalanalysisof訂ansgenicplantsshowedth

8、attherewerelittledifferencetochlorophyllcontentsandratioofchl日tochlb(chla／b1betweentheWTand訂ansgeneplantofArabidopsisThegrowthkineticsandchloropyllfluorescenceanalysisshowedretardedgrowthrateoftransgenicArabidopsisThus，t