东方粘虫微卫星富集文库的构建与遗传标记筛选的开题报告.docx

东方粘虫微卫星富集文库的构建与遗传标记筛选的开题报告

Topic:ConstructionofMicrosatelliteEnrichmentLibraryandGeneticMarkerScreeninginOrientalFruitMoth

Introduction:

Theorientalfruitmoth(OFM),Grapholitamolesta,isaseriouspestofstonefruitandothercropsworldwide.Managementofthispestreliesheavilyonpesticides,whichcanresultinthedevelopmentofresistanceandothernegativeenvironmentalimpacts.Therefore,alternativeapproachessuchasgenetics-basedcontrolstrategieshavebeenproposed.Microsatellites,alsoknownassimplesequencerepeats(SSRs),areidealmolecularmarkersforgeneticstudiesduetotheirhighvariability,co-dominantinheritance,andeaseofgenotyping.MicrosatelliteenrichmentlibrariesallowtheidentificationanddevelopmentofSSRmarkers,pavingthewayforstudyingthepopulationgenetics,ecology,andbiologicalcontrolofOFM.

Objectives:

1.ToconstructamicrosatelliteenrichmentlibraryfromOFMgenomicDNA.

2.ToscreenthemicrosatellitemarkersandselectthemostinformativeonesforfurtheruseingeneticstudiesofOFM.

Methods:

1.GenomicDNAextraction:GenomicDNAwillbeisolatedfromadultOFMindividualsusingastandardprotocol.

2.Constructionofmicrosatelliteenrichmentlibrary:Thelibrarywillbeconstructedusingaprotocolthatinvolvestheenrichmentofmicrosatelliterepeatsusingbiotin-labeledprobes,followedbyPCRamplificationandcloningoftheenrichedDNAfragments.

3.Microsatellitemarkerscreening:ThelibrarywillbescreenedusingPCRamplificationandsequencingoftheenrichedDNAfragments.TheresultingsequenceswillbeanalyzedforthepresenceofSSRsusingappropriatesoftware.PrimerpairswillbedesignedforamplifyingtheselectedSSRmarkers,andtheirpolymorphismandinformativenesswillbeevaluatedusingapanelofOFMindividuals.

Expectedoutcomes:

1.AmicrosatelliteenrichmentlibraryofOFMwillbeconstructedandevaluatedforitsefficiencyincapturingSSRs.

2.