30037.重組納豆激酶基因在雙子葉植物中的轉(zhuǎn)化與表達(dá)

1、湖北大學(xué)碩士學(xué)位論文重組納豆激酶基因在雙子葉植物中的轉(zhuǎn)化與表達(dá)姓名：張靜申請學(xué)位級別：碩士專業(yè)：微生物及生化藥學(xué)指導(dǎo)教師：楊艷燕20090501AbstractNattokinaseisastrongfibrinolyticenzymeinthetraditionalJapanesesoybeanfoodnattoTheenzymenotonlystronglyhydrolyzesthrombiinvivo，butalsoactives

2、plasminogentoplasminItisaproducedbyBacillussubtflisAttentionshavebeenpaidonitforitsadvantagesofsuperabilityofdissolvingthrombus，safety，longhalftimeperiodandbeingeffectivebyoraladministrationInthisresearch，nattokinasegene

3、betransformedintotobaccoandtomatoplantsbyAgrobacteriummediatedHereisthesummaryofworkshavebeendone：1TheNattokinasegenewasclonedintoexpressionvectorpCIAMBIA2300TherecombinantwasnamedastheplantexpressionvectorpCIAMBIA2300一N

4、attokinaseTheNattokinasehasa35s一5’sequenceasthepromoterintheupstream；anos3’sequenceastheterminatorinthedownstream2nenewvectorwasconstructedandnamedEHAl05pCIAMBIA2300一NattokinasebytransferringtheaboveconstructintoAgrobact

5、eriumtumefaciensstrainEHAl05usingthefreezeandthawmethod3Tobaccowasinfectedbyusingtheagrobacterium—mediatedleafdisctransformationPlantswereregeneratedonselectivemediawithKanamysinThepresenceoftheNattokinasegenewasconfirme

6、dbyPCRinplantssurvivingsectionsRTPCRresultsshowedthatNattokinasegenehasbeenexpressedatthetranscriptionlevel4Tomatowasinfectedbyusingtheagrobacterium—mediatedleafdisctransformationPlantswereregeneratedonselectivemediawith

7、KanamysinThepresenceoftheNattokinasegenewasconfirmedbyPCRinplantssurvivingsections5Proteinswereextractedfromthetransgeneicplant’SlevesandanalyzedbySDS—PAGETheresultssuggestedthattheNattokinasegenewasexpressedbytransgenei