鴕鳥微衛(wèi)星DNA的克隆、篩選與特性分析.pdf

1、中國農(nóng)業(yè)大學(xué)碩士學(xué)位論文鴕鳥微衛(wèi)星DNA的克隆、篩選與特性分析姓名：湯波申請學(xué)位級別：碩士專業(yè)：動物遺傳育種與繁殖指導(dǎo)教師：張勞李寧2002.6.1墅呈塑里里型壘塑塞墮：墮壅絲塑堡坌塑ABSTRACTMicrosatellitemarkersatewidelyusedinlinkagemapping，populationgeneticstudies，molecularevolutionandparentagetestinginmanya

2、griculturalspecies，whileonlyalimitednumberofostrich(Struthiocamelus)microsatelliteshavebeenisolatedThus，weconstructedtheostrichsmallinsertgenomiclibrariesofnon—enrichedandenridledformicrosatellitascontainingCArepeat111ee

3、nrichedmethodincludesligationof3001000bpenzymedigestedfragmentsandadaptors，affinitycaptureofmicrosatelliterepeatregionsusingbiotinylatedoligonucleotideattachedtostreptavdincoatedmagneticbeads，andPCRamplificationusingthe2

4、1metadaptorolignnucleotide∞primertoobtaindouble—strandedtargetedfragmentscontainingCArepeatWeisolated14clonescontainingCArepeatfrom3462clonesofnonenrichedlibrariesbyradioactivescreeningand499positiveclonesfrom603clonesof

5、enrichedlibrariesbyPCRscreeningUsingtheenrichmentprocedures，theproportionofclonescontainingCArepeatwasraised8270comparedwitllO4％inthenonenrichedlibrariesindicatinganenrichmentvalueover200foIdresultingin1esstimeconsumptio

6、nandleSScostforcloningThenumberofcompletesimpleCArepeatsinthesepositiveclonesrangedfrom5to29Theprimersfor94ofthesemicrosatellitesweredevelopedandusedtodetectpolymorphismswithmulti—PCRandgenomicscanning62ofthesemicrosatel

7、litelociexhibitedlengthpolymorphismsin94DNAsamplesextractedfromourostrichreferencepopulationsThenewpolymorphicmicrosatellitemarkerswehaveidentifiedandcharacterizedwillcontributetotheconstructionofanlinkagemapofostrichgen

8、ome，populationsgeneticsstudiesandevolutinnrelationsstudiesofostrichandrestavianspeciesThisapproachofconstructingandscreeningofmicrosatelliteenrichedlibrariesisalsoapplicableinthemicrosatelliteisolmionsofotherspeciesKeywo