24334.兩個野大麥cbls基因家族成員的研究

1、分類號Q75學校代碼10129UDC577學號08210032兩個野大麥CBLs基因家族成員的研究StudiesofTwoMembersofCBLsGeneFamilyfromHdeumbrevisubulatum申請人：吳廣宇學科門類：理學學科專業(yè)：生物化學與分子生物學研究方向：植物生物化學與分子生物學指導教師：李國婧教授李瑞芬研究員論文提交日期：二〇一一年六月StudiesofTwoMembersofCBLsGeneFamilyfr

2、omHdeumbrevisubulatumAbstractCBLs(calcineurinBlikeproteins)aclassofplantspecificCa2senswhichwereiginallyidentifiedinA.thalianawerenamedastheyaresimilartoCNBNCSinanimals.CBLsplayimptantrolesinplant’sspecificgrowthdevelopm

3、entprocessesstressresponseswithitsinteractionproteinCIPK(CBLinteractingproteinkinase).ResearchontheCBLsiscurrentlyfocusedinArabidopsisricebuttherewasverylittlestudyontheotherplants.Shtsubulatebarley(Hdeumbrevisubulatum(T

4、rin.)Link)isoneofthemostsalttoleranthalophytesinTriticeaewhichhasevolvedthemechanismofadaptationtosalinityduringlongtermofevolution.Itisvaluablefresearchapplication.CloningCBLsinShtsubulatebarleynotonlyshowstheeticalmean

5、ingsbutalsobenefitstopracticalapplicationprovidingpotentialcidategenesfimprovingstressestlerancebygeictransfmationinplant.HereweisolatedtwogenesinShtsubulatebarleynamethemHbCBL1HbCBL2.Weanalyzedtheresponseofthegenestodro

6、ughtstressessubcellularlocationtheirfunctionthemainprogressesareasfollows:1.ClonesequenceanalysisofHbCBLs:RTPCRwasusedtoclonefulllengthofHbCBL1HbCBL2.UsingsoftwareanalysisweknowthatbothHbCBL1HbCBL2hasCBLspeicificCa2bindi

7、ngsiteEFhsHbCBL1hasaNmyristoylationsiteatitsNterminaldomain.2.Fussionexpressvectswereconstructedtransfmedintoonionepidermalcellsbyparticlebombardmentrespectively.ThesevectswerealsotransfmedintoArabidopsisbyflaldip.Confoc

8、alobservationsshowedthatHbCBL1accumulatedinplasmamembraneaswellasinnucleusHbCBL2wasaccumulatedintonoplast.3.UsingNouthernblottoanalysistheexpressionofHbCBLswhileunderdroughtstess.WithoutanystressHbCBL1HbCBL2haveasmallamo

9、utofbackgroundespressionexpressioninrootsismestrongthanintheleaf.BythetreatmentofPEG6000mannitolespressionsofHbCBL1HbCBL2startedtodiminishastheprocessingtimewentby.4.IndertoexplethebiologicalfunctionofHbCBLsseveraloverex