大豆異黃酮代謝途徑在大腸桿菌中的構(gòu)建與表達(dá)

1、中國農(nóng)業(yè)大學(xué)碩士學(xué)位論文大豆異黃酮代謝途徑在大腸桿菌中的構(gòu)建與表達(dá)姓名：郝佳申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：農(nóng)產(chǎn)品加工與貯藏工程指導(dǎo)教師：陳尚武馬會(huì)勤20070601Soybeanisoflavoneshavemanygoodeffects∞hmshealthandtheresearchaboutithasbeenhottopicsinmany矗e№Abstractedfromsoybeanisthemainmethodofobtainingi

2、soflavoneAsplantsecondarymetabolitesisoflavones躺lowcontentinsoybeanandthegrowthcycleofsoybeanisrelativelylongSotheyieldofisoflavoneislimitedAsthedevelopmentofbiotechnologyandsoybean釅邶斌usingrecombinanttoproduceisoflavoneb

3、ygonetic鋤gi粥酬ngbecom瞄possibleIt’s童I踟methodtoobtainisoflavonelargelyandquicklyharmsimportantsi卿6∞∞eInthispapfr9soybeanleavesthathasbeenunderUVconditionsWaSusedasthestartingmaterialtoeOllSUoctfulllengthcDNAlibrazybySMARTte

4、chnologysixgenesofisoflavonoidbiosyntheticpathwaywerequicklyamplifiedbyPCRmethodwiththeonginallibrarycDNAdilution鷓templatesTheDNAsequencingindicatethecloneofgenesweresuccessfulEcoilexpressionvectorofeachgonew鴰constructed

5、andexpressedillEcoilBL21(DE3)straintheanalysisoftotalEcolisolubleproteinsbySDS—PAGEindicatethatallthegonespecificproteinswereavailableexceptC4HItalsoshowedthattheproteinsco∞W8584andgreaterthan78significantly011p005aboutC

6、HSinMS，whichmeansitissoybeanCHSThenthefivegeneswithoutc4hweretransformedtoEcoiLBL21(DE3)bytwoexpressionvectorseverygonehasitsownT7promoterandRBSneresultsofidentifyingexperimentsshowedthetwovectorscarryingmultigenecouldco

7、existsteadilylongenoughtoexpressionandfermentationTherecombinantba刪acarryingthewholeisoflavonoidbiosyntheticpathwaywmfermentedwithLtymsineassubstrateandIPTGas∞inducertherecombinantbacteria㈨produceanewisoflavonerelatedmet

8、aboliteshowedonHPLCanalysisprofile111eresultofMS2showedthefermentproductcontainsequol，whichisthesecondarymetaboliteofisoflavoneFurtherstudyofthenewproductisunderwayTheexperimentalresultsindicatedtheconstructionandexpress

9、ionoftbewholesoybeanisoflavonoidbiosyntheticpathwayinEcoilWaSsuccessfultherecombinantbacteriacanproduceanewisoflavoneItprovidesimportantfoundationforfurtherresearchoffindingnewresourceofisoflavonesKeywords：Soybeanisoflav