三種昆蟲病原線蟲Enolase基因的克隆、原核表達及功能研究.pdf

1、福建農(nóng)林大學碩士學位論文三種昆蟲病原線蟲Enolase基因的克隆、原核表達及功能研究姓名：高秋峰申請學位級別：碩士專業(yè)：農(nóng)藥學指導教師：吳珍泉邱德文20100401福建農(nóng)林大學2010屆碩士學位論文AbstractEntomopathogenienematodes(EPNs)isakindofimportantfactorinpestbiocontr01Nowadays，ithasbeenusedwidelyinagricultural

2、productionEnolaseisakeyenzymeinglycolyticpathwayHoweverrecentstudiesshowedthatenolasewasamultifunctionalprotein，andparticipatedintheinvasionofsomepathogensGenecloning，expressionandfunctionalresearchofenolasefromthreekind

3、sofEPNs，includedSteinernemafeltiae，SteinernemacarpocapsaeandHeterorhabditisindica，werecarriedout1UsingRTPCRandRACRtechnique，thefull—lengthenolaseeDNAfromthreeEPNswereclonedEnolasegenefrom＆feltiaeWas1453bpinlength，include

4、danopenreadingflame(oPt)of131lbp，encoding436aminoacidresidues；Theisoelectricpointandmolecularweightoftheproteinwerepredictedas567and4730982，respectively；GenBankaccession110HM067751Enolasegenefrom＆carpocapsaeWas1405bpinle

5、ngth，includedanopenreadingframe(oPt)of1311bp，encoding436aminoacidresidues；Theisoelectricpointandmolecularweightoftheproteinwerepredictedas559and4715461，respectively；GenBankaccessionnoHM067750Enolasegenefrom＆feltiaeWas161

6、5bpinlength，includedanopenreadingframe(ORE)of1305bp，encoding436aminoacidresidues；Theisoelectricpointandmolecularweightoftheproteinwergpredictedas543and4726863，respectively；GenBankaccessionnoHM067750Multiplesequencealignm

7、entindicatedthatenolasefromthreeEPNssharedmorethan75％identitywiththoseofothersixnematodespeciesattheaminoacidlevel，showinghighhomologyamongthemPhylogenetictreeindicatedthat11indicahastheclosestrelationshipwithCbriggsae，S

8、feltiaeandscarpocapsaehavetheclosestrelationshipeachother2ThreeenolasegeneswereclonedintopGEX6P一1toconstructGSTTagfusionexpressionvectors，andwereexpressedsuccessfullyinEscherichiacoli，respectivelyAndthenrecombinantprotei