小麥耐鹽突變體TaGSK1基因的分離和鑒定.pdf

1、河北師范大學(xué)碩士學(xué)位論文小麥耐鹽突變體TaGSK1基因的分離和鑒定姓名：徐濤申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：細(xì)胞生物學(xué)指導(dǎo)教師：沈銀柱黃占景20030601徐濤：小麥耐鹽突變體揚(yáng)GSKI基因的分離和箍定ABSTRACTOurpeviousstudiesshowedasfollows：firstlythefragmentofTaGSKlgenewasclonedandsequencedsuccessfullyTheaminoacidsequenc

2、eandthestructuresoftheproteinwereanalysedbyusingsoftwaresformolecularbiologyThecDNAsequenceofTaGSKlfrom力。iticumasetium￡wasregisteredatGenBankasanewgonewiththeaccessionnumberofAF525086Insuccession，usingDNArecombinanttechn

3、ique，arecombinantproteinexpressionveclor—pBV221gsk]andpBll21gsklof而G^張』wasconstructedbasingontheclonevector—pD—T23AtieranalysisandexpressionjnEcoliDH5oandBL2lof而G5KJcDNAasepcialexpressionproteinofmolecularweightof435KDca

4、nbeproducedandthequantityofexpressedapartinthemreachto8％and108％Throughanalyzingthesalttolerantcharacterofthetransfonnant，wefoundthatthesaltresistanceofthetransformantwashigherthanhostbacteriumInordertodistinguishthedifle

5、renceofTaGSKIbetweensaIt—stressresistantandsaltstresssensitiveontheDNAlevelweusedthesameprimersamplyfyandclonethefragmentsofTaGSK』DNAfromthegenomeofwheatlinesRH870649andH870634，ThesequenceresultsindicatedthatTaGSK』geneha

6、shighconservationandthedifiefenceofit’ssaltresistanceexpressedmainlyonthetranscriptionandtranslationlevelhoweverontheDNAlevelthereisnoanydifierence1thasthreecopiesinthegenomethatwastestifiedbySouthernBlotItwasrarereporte

7、dinourcountrythatmatureembryoderivedcalljfromWheatasthematerialsweretransfcIrmedwithAgrobacteriumtunefaciensWestudiedtheiraGSKI’stransformationofWheat(Triticumasetium￡1respectivelythroughtwomethod：Agrobacteriumtumefacien

8、s—mediatedtransformationandparticlebombardmeritusingthesamematerialsofmatureembryo—derivedcallifromsaltstresssensitiveWheatIinesandtilebinaryplantvectorpBll21gsklTransgeniccallusofWheatwereproduced，usingKnaandNaCIforsele

9、ctionThecallusweredemonstratedpositivewhengenomicDNAoftransgeniccalluswasidentifiedbythemethodofPCR，usingtwoterminationsequenceofNPT—IIgeneasprimersTransgeniccallusofWheatcangrowprosperousin05％NaCIhoweverthecontrollinesa

10、lmostallchangedbrownevendiedOurpilotstudyindicatedthatTaG脒，genecanenhalacethetoleranceofsaltstresssensitiveWheatlinesInadditionourstud)showedthatthetwotransfonnationmethodshaveexcellenceandshortcomingofthemselvesandthera

11、testransformationjsrelativewiththegenotypecloselyTheseresultshadimportantsignificancetesearchdeeplytheexpressioncontrolandprotienfuncfionofTaGSK』toopenoutthemechanismofsalttoleranceinWheatandtoexploitandusethetransformat