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苏云金芽胞杆菌新型杀虫基因的发掘与活性分析.docx

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苏云金芽胞杆菌新型杀虫基因的发掘与活性分析 Title:DiscoveryandActivityAnalysisofaNovelInsecticidalGenefromSerratiamarcescens Abstract: Inrecentyears,thecontrolofinsectpestshasbecomeincreasinglychallengingduetothedevelopmentofinsectresistanceandthenegativeimpactofchemicalinsecticidesontheenvironment.Inlightofthis,thereisagrowinginterestindiscoveringnovelandeco-friendlyapproachesforpestcontrol.ThisstudyaimstoexploreandanalyzethepotentialofanewlydiscoveredinsecticidalgenefromSerratiamarcescens,abacteriumknownforitsinsecticidalproperties. Introduction: SerratiamarcescensisaGram-negativebacteriumthathasgainedsignificantattentionasapotentialbiocontrolagentagainstvariousinsectpests.Itsabilitytoproducesecondarymetaboliteswithinsecticidalactivitymakesitapromisingcandidateforthedevelopmentofbiopesticides.Inthisstudy,wefocusonthediscoveryandcharacterizationofanovelinsecticidalgenefromS.marcescens,referredtoasSYP-1. Methods: 1.BacterialStrainandCultureConditions:S.marcescensstrainXwasisolatedfromsoilcollectednearacropfieldandusedthroughoutthestudy.ThestrainwasculturedinLuria-Bertani(LB)mediumunderoptimumgrowthconditions. 2.GenomicDNAExtraction:GenomicDNAofS.marcescensstrainXwasextractedusingacommercialkitfollowingthemanufacturer'sinstructions. 3.CloningandExpressionofSYP-1:TheSYP-1genewasamplifiedusingpolymerasechainreaction(PCR)withspecificprimersandclonedintoasuitableexpressionvector.TherecombinantplasmidwasthentransformedintoEscherichiacoliforproteinexpression. 4.PurificationandCharacterizationofSYP-1:TherecombinantSYP-1proteinwaspurifiedusingaffinitychromatography,anditsmolecularweightwasdeterminedthroughsodiumdodecylsulfate-polyacrylamidegelelectrophoresis(SDS-PAGE).Theprotein'sinsecticidalactivitywasassessedagainstkeyinsectpeststhroughbioassays. 5.ModeofActionAnalysis:ThemodeofactionofSYP-1wasinvestigatedthroughobservationsofhistologicalchangesinthetargetpestsusinglightandelectronmicroscopytechniques. Results: 1.CloningandExpressionofSYP-1:TheSYP-1genewassuccessfullyamplifiedandclonedintotheex