大豆異黃酮代謝途徑在大腸桿菌中的構建與表達

1、中國農(nóng)業(yè)大學碩士學位論文大豆異黃酮代謝途徑在大腸桿菌中的構建與表達姓名：郝佳申請學位級別：碩士專業(yè)：農(nóng)產(chǎn)品加工與貯藏工程指導教師：陳尚武馬會勤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