Molecular Cloning, Expression and Molecular Modeling of Chemosensory Protein from Spodoptera litura and Its Binding Properties with Rhodojaponin III 英文参考文献.docVIP

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Molecular Cloning, Expression and Molecular Modeling of Chemosensory Protein from Spodoptera litura and Its Binding Properties with Rhodojaponin III 英文参考文献.doc

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Molecular Cloning, Expression and Molecular Modeling of Chemosensory Protein from Spodoptera litura and Its Binding Properties with Rhodojaponin III 英文参考文献

MolecularCloning,ExpressionandMolecularModeling ofChemosensoryProteinfromSpodopteralituraandIts BindingPropertieswithRhodojaponinIII YanboZhang,XiaolinDong,JinxiangLiu¤,MeiyingHu*,GuohuaZhong*,PengGeng,XinYi Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People’s RepublicofChina Abstract Insectsstimulatespecificbehaviorsbythecorrectrecognitionofthechemicalsintheexternalenvironment.Rhodojaponin IIIisabotanicalgrayanoidditerpenidovipositiondeterrentisolatedfromRhododendronmolle.Inthisstudyweaimedto determinewhethertheCSPsinvolvedintherecognitionofRhodojaponinIII.Afull-lengthcDNAencodingchemosensory proteinwasisolatedfromtheantennaeofSpodopteralituraFabricius(CSPSlit,GenBankAccessionNo.DQ007458).Thefull- lengthcDNAofNlFoxAis1789bpandhasanopenreadingframe(ORF)of473bp,encodingaproteinof126aminoacids, Northern blot analysis revealed that CSPSlit mRNA was mainly expressed in the antennae, legs, wings and female abdomens.Athree-dimensionalmodelofCSPSlitwasconstructedusinghomologymodelingmethod,anditsreliabilitywas evaluated.TheactivesiteofCSPSlitwascalculatedusingCDOCKERprogramindicatedthattheTyr24,Ile45,Leu49,Thr64, Leu68, Trp79 and Leu82 were responsible ligand-binding active site on identifying Rhodojaponin III in the CSPSlit. The recombinant CSPSlit protein was expressed in Escherichia coli and purified using single-step Ni-NTA affinity chromatography.FluorescenceemissionspectrarevealedthattheCSPSlitproteinhadsignificantaffinitytorhodojaponin III.TheseresultsmeanthatCSPSlitiscriticalforinsectsidentifytheRhodojaponinIII. Citation:ZhangY,DongX,LiuJ,HuM,ZhongG,etal.(2012)MolecularCloning,ExpressionandMolecularModelingofChemosensoryProteinfromSpodoptera lituraandItsBindingPropertieswithRhodojaponinIII.PLoSONE7(10):e47611.doi:10.1371/journal.pone.0047611 Editor:EugeneA.Permyakov,RussianAcademyofSciences,InstituteforBiologicalInstrumentation,RussianFederation Receive