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《三疣梭子蟹微衛(wèi)星文庫的構(gòu)建及多態(tài)性位點篩選》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、●◆上海海洋大學(xué)碩士學(xué)位論文MicrosatellitelibraryConstructionandpolymorphiclociscreeningfortheswimmingcrab，PortunustrituberculatusABSTRACTmswimmingcrab(Portunustrituberculatus)isanimportantfisheryresource，widelydistributedincoastalareasofChina．11圮swimmingcrabwasregarededasoneofthemarineaquaculturespeciesfrom198
2、1．Ithasbecomeoneofthemainaquaculturespeciesduetoitsconsiderableeconomicvalue，highediblenutritionalvalueandfastgrowingspeed．Becauseofvariousreasons，theresourcesofPtrituberculatusdroppedsharply，defenseabilitydeclinedandnaturalgermplasmresourcesdegradedseriouslyinrecentyears．Inordertorestoreandimprov
3、ethequalityoftheparentcrabandcrabseed，keepthehealthydevelopmentofaquaculture，avoiddecrementofwildgermplasmresources，andrealizesustainableutilizationofthenaturalresources，researchonthegeneticgermplasmresourcesofP．trituberculatusisneededurgently．Therefore，developingasetofeffectiveandreasonablemolecu
4、largeneticmarkersisnecessaryforgeneticgermplasmresourcesconservationresearchforthisspecies．MicrosatelliteDNAlibraryoftheswimmingcrabWasestablishedusingthemagneticbeadhybridizationenrichmentmethods．T0talgenomicDNAWasextractedanddigested、析tllrestrictionenzymeSau3AI．Fragmentsinrangeof200--1000bpwerea
5、garose-gelpurifiedandligated、析tllspecialadaptor,andthecombinationsWashybridized、Ⅳitllbiotin—labelledmicrosatelliteprobe(CAh2，(GA)i2，(TGC)s．Byusingthehighbindingaffinityofbiotintostreptavidin—coatedmagneticbeads，single-strandedDNAcontainingtheselectedmicrosatellitesequenceswerecaptured．ThetargetedD
6、NAWasused鷦thetemplateforPCRamplificationusingtheadapter嬲primers；andtheproductswerethencloneddirectly,screenedbybacteriumliquidPCRmethodsubsequently．Ofthe60positiveclones，42microsatellitelociwereacquired，whichwerefurthercategorizedintoperfect(73．8％)，imperfect(21．4％)，andcompound(4．8％)types．Amongthem
7、icrosatelliterepeats，10,--30repeatsweremostabundant．III●上海海洋大學(xué)碩士學(xué)位論文ByusingPremierPrimer5．0，wedesigned31pairsofmicrosatelliteprimers，amongwhich16pairscanamplifyexpectedproduct．These16pairsofprimerswerefurtherused
