馬氏珠母貝啟動子的克隆和轉(zhuǎn)基因技術(shù)的探索.pdf

1、海南大學(xué)碩士學(xué)位論文馬氏珠母貝啟動子的克隆和轉(zhuǎn)基因技術(shù)的探索姓名：黃海燕申請學(xué)位級別：碩士專業(yè)：水產(chǎn)養(yǎng)殖學(xué)指導(dǎo)教師：王愛民石耀華20090501AbstracPinctadamartensii(Dunker)isoneoftheprincipalcommercialpearloystersinmariculture，thecultureofwhichhaslatelybeenapillarindustryinsomeareasalong

2、thesouthcoastofChinaHoweverabigbunchofnaturalresourceshavebeenfishedtoapointofextinctionitishightimetoimprovethequalityaswellasquantityofaquaticanimalsbyvirtueofmodembiotechnologyBasedon只martensiiasmainexperimentalmateri

3、al，thestudyhadbeendoneinthefollowingtwoways：1PromotercloneTAILPCRtechniquewasemployedtoamplifythepartofknown尸martensiieDNAsequencefromelongationfactor1一aandS9proteingeneof40SribosomalsubunitAbout1200bpoligonucleotidesobt

4、ainedfromthe5’endofstartcodonWasclonedandsequencedsoonafterTwonetworksoftware一‘NeuralNetworkPromoterPrediction’’and‘‘TheMarkovChainPromoterPredictionServer’—hadbeenutilizedinforecastingpromotershowingthatthepossibilityof

5、beingapromoterfortheamplifiedsequenceofelongationfactor1一awas33％，whereasthelikelihoodWasabove80％byamplifiedsequencefromS9proteingenein40Sribosomalsubunit2EstablishmentofgenetransfermethodTheplasmidwithGFPWasinjectedintot

6、hegonadofmalepearloysterswhichwerethenputtofertilizationItWasdifficulttotellifgenetransfersucceededornotbecausebothexperimentalgroupandcontrolgroupwereobservedtogiveadelicatefluorescenceinmicroscopeTheextractedgenomiclar

7、vaeDNAWasculturedformorethan48hoursandlatertestedbyGFPspecificprimerinPCRThepositiveresultrevealedthatexogenousplasmidmighthavealreadyenteredintothelarvaeNevertheless，GFPhadnotbeenincorporatedwiththechromosomeofreceptorE