抗性和敏感棉鈴蟲CYP6B7基因的克隆及不同發(fā)育階段的表達研究.pdf

1、中國農(nóng)業(yè)大學(xué)碩士學(xué)位論文抗性和敏感棉鈴蟲CYP6B7基因的克隆及不同發(fā)育階段的表達研究姓名：張衡申請學(xué)位級別：碩士專業(yè)：農(nóng)藥學(xué)指導(dǎo)教師：邱立紅20070601中國農(nóng)業(yè)大學(xué)碩士學(xué)位論文AbstractAbstractCouonbollwormHelicoverpaamigerafHabner)isaseriouseconomicinsectpestinagricultureallovertheworldSeriousresistanceo

2、fHarmigeratomanykindsofinsecticidesincludingpyrethroidshasoccurredCytochromeP450smediatedenhancedmetabolicactivitywasprovedtobcamajormechanismofHarmigeraresistanttopyrethroidsOverexpressionofP450sgece，substitutionofamino

3、acidsandjointactionofoverexpressionandaminoacidsubstitutionweresuggestedrelevanttothemsistanceStudiesoncytochromeP450genewouldcontributetobetterunderstandingthemolecularmechanismof丘armigeraresistance笛wellasprovidingtheor

4、eticalbasistomsislancePflⅢtgement磺HarmigeraandrationalapplicationofpyrethroidsCYP687genomieDNAsequencesfromⅡ1efanvaleraterusistancestrain(HDFRstrain)andthesusceptiblestrain(HDSstrain)wereobtainedfromthemidgutgenomicDNAof

5、thesixthinstarlarvaeof讎cottonbollwormbyusingapairofprimersspecifictoCYP687(GeneBankNoAF031468)TheresultshowedthatthesequenceofHDFRandHDSstrainwas1840and1842bpinlen舛hrespectivelybothhavinganopenreadingflameof1515nucleotid

6、esencodingaproteinof504aminoacidresiduescontaining325bpand32rapintronsinlengthrespectivelyComparingwithHDSstraintheidentitiesofnuelcotidesandaminoacidsofsequencefromHDFRstrainwere9924％and9940％respectivelyTherewe∞llmutati

7、vesitesinnueleotidesofcodingregionwhichresultedin3substitutionofdeducedaminoacidsinHDFRstrailLMoreovertherewere3mutativesitesintheintrothincluding2nuelcotidesmissingTheexpressionofCYP687mRNAatdifferentlifestagesofHarmige

8、rawasde￡ectedbyusingNorthernblottingTheresultsindicatedthatCYP687wasonlyexpressedinthelarvalstageofH∞r(nóng)aigerabutnotinthepupaeandadultsMoreovertheexpressionlevelincreasedasthelarvaedevelopedTheconservedregionhamebindingsig

9、naturemotifandtheconservedIhelixinthealignmentofsomeinsectP450proteinsprobablyinvolvedinresistancefromGaneBankservedasaguideforthesynthesisofdegenerateoligonueleotideprimersPrimerswereusedinthereveP3etranscriptionpolymer

10、isechainreaction(RTPCR)ofRNAfromthemidgutRNAofthesixthinstarlarvaeofHarmigeraresistanttofenvalernteTheeDNAfragmentssizedasdesignedwc他obtainedbutthesequencesofthesefragmentsshowedhighidentitieswithP450sequencesintheGaneBa